ResearchSaves Spring/Summer

Climbingfor Silas

Spring/Summer 2013

S AVESS AVES

FOUNDATION FOR BIOMEDICAL RESEARCHAWARDS THREE SCHOLARSHIPS TO OUTSTANDING CAL POLY POMONA PRE-VETERINARY STUDENTS

NAVY DOLPHINS ARE SAFEGUARDINGOUR NATIONAL SECURITY — ANDADVANCING HUMAN MEDICINE

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Climbingfor Silas

RESEARCHS AVESSpring/Summer 2013

FEATURES3 Welcome

4 Climbing for Silas© The Jackson Laboratory

4 Climbing for Silas© The Jackson Laboratory

8 The Ethic of Care© Oregon State University

11 A Healthy Bond: By Improving Pain Treatment,Therapy in Dogs, Research Offers MedicalInsight for Humans© Kansas State University

12 Fats and Flies© National Institutes of Health

16 Tackling Diabetes© Oklahoma State University

18 How Bees Decide What to Be© Johns Hopkins University

20 Tackling a Brain Tumor Deadly to Pups and People© Virginia Tech

23 Bay Watch© Tufts University

26 Body Bacteria Exploring the Skin’s Microbial Metropolis© National Institutes of Health

31 Hopkins Researchers Solve Key Part of Old Mystery in Generating Muscle Mass© Johns Hopkins University

32 Cancer Collaboration Could Someday Help Dogs and their Humans© Princeton University

34 Tackling a Fungal Disease© Oklahoma State University

36 Black Death Threat© University of North Carolina

38 Foundation for Biomedical Research AwardsThree Scholarships to Outstanding Cal PolyPomona Pre-Veterinary Students © Foundation for Biomedical Research

40 The Mystery of an Antıcancer Mechanism© University of Rochester

44 For Janice: Legacy of a Short Life© National Institutes of Health

Editor-in-Chief: Michael StebbinsResearchSaves™ is a registered trademarkof the Foundation for Biomedical Research818 Connecticut Avenue, NWSuite 900Washington, DC 20006

All articles are reprinted in their entiretywith the expressed written consent ofthe authors and their associated institutions. Each article and photo isprotected under U.S. copyright law andremains the sole property of the authorsand their respective institutions.

© ResearchSaves.org. 2013 All Rights Reserved

A toddler's devastating diseasespurs a Maine boy to use hisown legs and lungs to helpthose who can't.

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For article submissions & advertising, please email FBR at [email protected].

ResearchSaves is a semi-annual publication. The annual subscription price is $39 and includesshipping and handling. Complimentary issues are available to K-12 teachers, thanks in largemeasure to the generous sponsorships granted from individual biomedical researchers.

Climbing for Silas

11 A Healthy Bond. By improving pain treatment and therapy in dogs, a researcher is offering medical insights for humans.

16 Tackling Diabetes. A team ofresearchers hope that research withanimals can lead to help for humans.

23 Bay Watch. Navy dolphins aresafeguarding our national security, and advancing human medicine at thesame time.

32 Cancer Collaboration CouldSomeday Help Dogs and TheirHumans.

Climbing for Silas

Spring/Summer 2013 ResearchSaves.org 3

Greetings! Welcome to another issue of ResearchSaves.Once again, we have a lot of great storiesabout kids, animals and families. Thecommon thread that weaves throughout allthe stories is how biomedical research isallowing them to live happier, healthier lives.

Because of the great response we had to last issue's Liviya's Story, thismonth's cover story also features an amazing kid with an inspiring story.This time, though, the kid is raising money and awareness for a diseaseaffecting someone else. When Gus La Cosse, a 10 year-old in Maine,heard about kids with SMARD (spinal muscular atrophy with respiratorydistress), he knew had to do something to help out. And the rest, well,you'll have to read the story to find out more. It begins on page 4.

In this issue, we also have interesting profiles of some amazingresearchers at the National Institutes of Health, studying everythingfrom fats in flies to germs in mice to cancer in naked mole rats.

Have a comment about what you are reading in this issue? Send us aletter-to-the-editor at [email protected]. We'd love to include yourthoughts and ideas on how to improve the magazine, as well as hearyour comments about this issue's stories.

And last but not least, we're printing the winners of our 2012 AnimalResearch Essay Contest, in partnership with Cal Poly Pomona. Thethree winners wrote three very different takes on the subject of animalresearch. I think you'll find each of them interesting in their own right.

That's all for this issue. See you in the Fall!

Frankie TrullPresident

Nahla Al BassamAdministrative Assistant

Hometown:Baghdad, Iraq

Joined FBR:2008

Liz HodgeDirector, Mediaand MarketingCommunications

Hometown:Hamilton, MA

Joined FBR:2008

Paul McKellipsExecutive Vice President

Hometown:Neenah, WI

Joined FBR:2007

Cherie ProctorDirector ofDevelopment

Hometown:Quincy, MA

Joined FBR:2002

Michael StebbinsDirector ofResearch andEditor-in-Chief

Hometown:Silver Creek, NY

Joined FBR:1999

The FBR Team

4 ResearchSaves.org Spring/Summer 2013

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Gus La Casse did some math.

What he concluded was that if each of25 mountain treks could attract pledges,that money would provide a tremendousboost for SMARD research at TheJackson Laboratory and, he hoped, help find a cure.“My mom works at The Jackson Lab, and one day she came home andtold me about SMARD,” 10-year-old Gus La Casse said in July. “It’s prettyrare, but there are quite a few kids in America and kids all over the worldwho have SMARD. And nobody has a cure.”

That insight distressed Gus, who is into climbing and hiking and whorealizes that kids with SMARD will never do either. He decided he wouldspend much of his summer, in effect, doing that for them, while seekingpledges to fund research. It was, Gus says, a “good summer vacationdeed.”

Most people have never heard of SMARD, an acronym for spinalmuscular atrophy with respiratory distress, which Jackson LaboratoryAssociate Professor Greg Cox, Ph.D., describes as “the rarest of the rare”in terms of geneticbased, neuromuscular degenerative diseases.

“Until recently, I had never heard of it, nor have most physicians,” saysCox. “It’s a very debilitating infantile disease. Most of these children don’tlive more than two or three years. SMARD is a very early onset motorneuron disease. Motor neurons are the neurons in your spinal cord thatextend into every muscle in your body. When these motor neurons getsick—and with SMARD, they actually die—there’s no way to triggervoluntary muscle response, which includes breathing and swallowing. It’sa recessive disease. You have to have two parents who carry the mutationto have a fear of a child developing SMARD.”

Regrettably, Cox notes, he can count on the fingers of one hand thenumber of research laboratories worldwide that are exploring thebiomolecular complexities of SMARD. Researchers in those labs make useof a mouse model of SMARD provided through The Jackson Laboratory.

It was Greg Cox who discovered the genetic mutation that causes thiscurrently incurable disease that kills infants and toddlers through muscleatrophy and paralysis. While SMARD affects fewer than 1,000 Americanchildren at any given time, each case is devastating for the f amiliesinvolved. ➤


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Climbing for Silasfor Silas– Photography by FRANÇOISE GERVAIS

Among those families are the parents and grandparents of SilasWerner. As a newborn, Silas baffled a small army of Pittsburgh,Pa., pediatricians before he was finally diagnosed with SMARD,at age three months. After an original diagnosis of botulism anda prognosis of full recovery, agonizing months of hospitalizationfinally revealed a diagnosis of SMARD. His parents weredevastated.

When Gus, whose parents Renée and Joe both work at theLaboratory, first heard of Silas and SMARD, he was moved to act.

“I do a lot of things that involve movement, like hiking andclimbing,” he says. “So I decided I would raise money by doingsomething for these young kids with SMARD that they will neverbe able to do, like go hiking. In hiking, you have to breathe a lot,and these SMARD kids will never breathe on their own.”

Over his summer break from grade school, Gus trekked up anddown 25 mountain trails, most of them within Acadia NationalPark on Mount Desert Island. His final climb on July 31 was asteep ascent to the summit of Mount Katahdin, Maine’s highestpeak at 5,268 feet. It was five miles straight up, and five milesstraight down. Among those joining him in the trek was GregCox, who joined Gus on many of his climbs.

“When people ask me why they should contribute to Climb forSilas, I tell them because kids all over the world have SMARD,and there’s no cure.”

“He doesn’t get tired,” Cox says of Gus’ ambition. Apparently,neither does Greg Cox, according to his wife.

“The last couple of weekends have been very hot here on MountDesert Island,” Kathy Cox wrote in late July on the Facebook siteGus created to promote the project. “But Gus, Renée, Greg andteam kept hiking for SMARD families. We are so proud of you.”

Gus set an ambitious goal for his fundraising in hopes that GregCox can hire a postdoctoral researcher to focus on gaining abetter understanding the disease. To spread the word about hisproject, he was interviewed on a morning talk radio show andwrote an op-ed that he sent to the editorial pages of dailynewspapers throughout Maine.

“When people ask me why they should contribute to Climb forSilas, I tell them because kids all over the world have SMARD,and there’s no cure,” he wrote. “It’s important that people likeGreg Cox find a cure.”

Gus has never met Silas, but plans to. He’s been communicatingwith Lisa and John Werner, Silas’s parents, by e-mail, as well asother SMARD families he’s tracked down online, including afamily in England.

“There’s another kid, Dakin, who lives in Texas,” Gus says. “He’sfour years old, which is quite old for a kid with SMARD. I alsoheard that there’s a SMARD kid in California who is 18, which isvery unusual.”

The dust has settled on his summer of climbing, and Gus is backinto his usual school routine. But he continues to work toward hisfundraising goals to support research and discovery on behalf ofa far-flung community of children and parents to whom he is nowfirmly connected.

Silas Werner is beloved for his sweet disposition.

Silas Werner, the boy whoinspired Gus La Casse to climb and raise money forSMARD research, is now 2.

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Despite his condition, his parents say that Silas radiates joythrough his happy smile. Speaking haltingly through a ventilator,Silas has now mastered “mama” and “papa,” much to thedelight of his parents. Silas’ nurses describe him as “having thesweetest disposition of any baby we’ve ever met.”

Nonetheless, Silas requires the around-the-clock care providedby his parents and by visiting nurses. Lisa and John Werner havemastered what Lisa terms “special needs boot camp” to meetSilas’s needs.

The Werners are part of the close network of SMARD parentswho now have newfound hope for the future. They recentlylearned that The Jackson Laboratory has a laboratory makingstrides in SMARD research. They are actively raising funds tosupport the research and, hopefully, advance the understandingof the disease to the point that doctors can one day help Silasand his peers.

Contributions to support the fundraising efforts of Gus, Renéeand Joe can be made by visiting their webpage.

When people ask me why theyshould contribute to Climb forSilas, I tell them because kids allover the world have SMARD,and there’s no cure,” Gus wrote.“It’s important that people likeGreg Cox find a cure.”

The three rats snoozing inCage 57 don’t know it, butthey could someday help savethousands of human lives.Snuggled in their EcoFresh bedding, the rodents aredigesting a meal that may hold clues to preventingcolon cancer, the second-leading cause of cancerdeaths in the United States. On their cage, equippedwith HEPA air-purification filters and precisiontemperature controls, hangs a blue index card labeled“Special Diet,” on which a researcher has scrawled“Bruss” in black felt pen. The scrawl is short for Brusselssprouts, those oft-disparaged veggies resembling tinycabbages that are loaded with promising cancer-prevention compounds such as sulphoraphane.

To the rats, however, the pale-green pellets in their foodtray (Mix AIN93 from Research Diets Inc., with sproutsadded) are just dinner. That dichotomy — the rats’ bodily,mental and social needs (rodents are housed with“buddies” for company and “crawl tunnels” forenrichment) versus the precise methods of science —requires researchers to walk a tightrope, alwaysbalancing the pressing questions of medicine, forexample, against the welfare of animals. The results arekey to curing devastating diseases like ALS or Alzheimer’s.

Oregon State University, with 600,000 research andteaching animals (mostly fish and other aquatic species)at 30-plus sites across the state, is balancing thoseinterests exceedingly well. That is the judgment of theAssociation for Assessment and Accreditation ofLaboratory Animal Care International (AAALAC), whichin March gave a glowing report after an extensiveaccreditation study (see “High Grades for AnimalCare” – http://oregonstate.edu/terra/2012/10/high-grades-for-animal-care/). Oregon State is the 19thamong the nation’s 71 land grant universities to earnfull-campus AAALAC accreditation.

The snow-white, sprout-eating rats in OSU’s state-of-the-art rodent facility are just one among 400 vertebratespecies that populate the university’s labs, barns,aquariums, ranches and hatcheries. Zebrafish,steelhead, beef cattle, garter snakes, rainbow trout,dairy cows, yellow- and red-legged frogs, copper andcanary rockfish, lambs, koi, swine, salmon smolts andllamas are among the half-million-plus warm- and cold-blooded creatures that help educate OSU’s students,improve health (both human and animal), protectecosystems, guide resource management, bolster localeconomies and engage the public.

Every last one of these creatures, from the 2-inch troutfingerling to the 2,000-pound Hereford bull, is theresponsibility of Dr. Helen Diggs. If you don’t have anelectronic key card, you must knock at a security door togain admittance to her building on the west end ofcampus, the base from which Oregon State’s attendingveterinarian oversees her vast menagerie. With thewelfare of thousands of animals on her mind, she isquick to question, slow to trust (or, as she likes to say, “Itrust but verify”). It’s a hyper-vigilance honed over 25

BY LEE SHERMAN

“All who care for, use,or produce animalsfor research, testingor teaching mustassume responsibilityfor their well-being.”

Guide for the Care andUse of LaboratoryAnimals, 2011, TheNational Research Council.

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years in the field, some of those years at UC Berkeley where Diggsendured threats from animal-rights activists and had to beescorted to her car by security guards.

Tacos and M&Ms“This is my morning health-status report,” says Diggs, pointing toa spreadsheet on her computer monitor. “Every day, every animal-facility manager checks in with me. Here’s Chad Mueller at the agexperiment station out in Union. Here’s Rob Chitwood at the fishperformance lab over by the golf course. Here’s the Linus Paulingbuilding. The Oregon hatchery. The Horse Center. Wherever I am,I can open up this online report and see what’s happening.”

Doctor at the TopSo when a hamster is lethargic or a horse is lame, she’s on it. Butpractices haven’t always been so rigorous in the world of animalresearch. Diggs has been in the field long enough to have seenthe transformation.

“In the decades before the 1980s, some universities were notcaring for their animals as well as they should,” says Diggs, whohas overseen research animals since 1985. “The facilities smelledgnarly. There were wooden floors with urine stains, poortemperature control. Regulations weren’t being enforced. No onewas watching.”

This laxity was not just a problem for the animals. It was also aproblem for the science.

“Researchers weren’t able to repeat their results. If I’m keeping myrats in a closet and feeding them oatmeal for breakfast, while yourrats are getting leftover tacos or pancakes from the studentlounge, we can’t validate our findings.”

Adds Steve Durkee, another of the university’s leading research-animal watchdogs: “If rats in one study are getting cereal whilethose in a second study are getting oranges and M&Ms, you can’tcompare the results of the studies. By standardizing andharmonizing how animals are cared for, you create consistencyacross labs and institutions.” In fact, he notes, prestigiousacademic journals publish only findings that document the higheststandards of animal care.

That’s why Diggs’ job has teeth. Sharp ones.

“I can shut a program down,” she says. “I’ve never had to do ithere. But two times at other universities, I had to actually shutsomeone down and lock the door. If I have to go in and have aconversation with someone about their animal work, they’ll listento me. It’s a big deal.”

Even though the U.S. Public Health Service mandated in the early‘70s that all animal research institutions hire an attending vet, thetop docs didn’t have any real enforcement power until the mid-’80s. That’s when the National Institutes of Health and the U.S.Department of Agriculture cinched up the rules for labs gettingfederal research dollars.

“Attending vets had no real authority back in the early days,”explains Diggs, who reports to Rick Spinrad, vice president ofresearch at Oregon State. “Some didn’t even have keys to theanimal facilities. You need someone who’s minding the store, notjust a figurehead.”

Minding the StoreBob Murray waves his key card in front of a laser-triggered securitypanel in the $62.5 million, 1-year-old Linus Pauling ScienceCenter, which houses the Department of Chemistry as well as theLinus Pauling Institute. The elevator opens, and he steps inside.One floor down, he flashes his card again, clicking open anelectronic steel door into a small anteroom, where he slips on agauzy yellow “isolation gown” and a pair of puffy blue booties.

For a third time, Murray brandishes his key card, unlocking yetanother heavy door. He enters the inner sanctum of OregonState’s gleaming “vivarium” — the small-mammal equivalent ofan aquarium or a terrarium — where hundreds of rats andthousands of mice live, as well as a few hamsters. Not one ofthese furry beings can get a sniffle or a sore toe without Murrayknowing about it.

“The animals are checked at least twice a day, 24-7,” says Murray,who clocked 35 years in the field, working at the New EnglandPrimate Center, Walter Reed Army Medical Center, Letterman ArmyMedical Center, UC Berkeley and Genentech before coming toOSU last year to manage the lab-animal facilities. “We watch forchanges in gait or overall appearance — does the animal’s coatlook scruffy? How is the animal’s appetite and hydration? We lookfor lethargy, weight loss, tumors. Any health problems we reportimmediately to Dr. Diggs and the researchers.”

The vivarium maintainsstrict controls ontemperature and otherfactors in the animals’environment.

Murray’s dad worked for the Society for Prevention of Cruelty toAnimals in Boston for nearly 30 years. So worrying about animalwelfare is practically in his genes. He takes pride in the life-savingresearch he has observed over the years, like the groundbreakingHerceptin research at Genentech that is being used to treatthousands of women with breast cancer and the malaria vaccineresearch at Walter Reed. Still, it’s the health and comfort of thewhiskered rodents that gets him out of bed every morning at 5o’clock and keeps him running as he oversees his team of highlytrained, certified animal technicians.

“I believe strongly in the value of the research we do here, but I’mnot a researcher,” Murray says, surveying his domain with thediscerning gaze of a seasoned professional. “I’m into animal care.”

If Murray were to take you through the 8,000-square-foot facilitywhere researchers investigate the links between nutrients andhuman health, the first thing you would notice is an obsession withcleanliness. The giant Steris cage washer (which he calls “theheartbeat of the whole facility”) sanitizes racks of cages in twocycles of 180-degree, pressurized water — and that’s after thecages have been blasted with detergent and rinsed in acid.Everywhere you look, technicians and student workers are preppingcages for incoming animals or plying mops on floors that alreadylook immaculate. Viruses and bacteria that could sicken theanimals and compromise the research don’t stand a chance.

The next thing you would notice is the attention to precision.Automated lighting simulates 12 hours of day, 12 hours of night.Electronic monitors maintain a 68- to 72-degree temperaturerange. An alarm alerts the staff if temperatures fall outside therange by even 1-degree Fahrenheit. There are ventilation ➤


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tubes, fume hoods, stainless-steel work stations illuminated withstretchable spotlights. Every last facet of the facility is designed toprotect the health and welfare of all its mammalian inhabitants,human as well as rodent.

Not until you reached the bosom of the vivarium would youcome upon the rodents. The Brussels sprout-eating residents ofthe “rat room” were born and raised at an Indiana-basedresearch-animal supply company called Harlan Laboratories,arriving at OSU in ventilated crates via UPS. Firms like Harlan,along with Charles River Labs, Jackson Labs and dozens of otherscomprise a global mega-industry in the service of science. Allmust adhere to the same stringent federal requirements thatguide OSU’s animal-care personnel.

Diets supplemented with cancer-fighting compounds are underinvestigation in the Linus Pauling Science Center.

In the rat room where Rod Dashwood and other researchers inthe LPI Cancer Chemoprotection Unit are looking for evidencethat cruciferous vegetables like Brussels sprouts and broccolisprouts can block the formation of colon tumors, dozens of clear-plastic cages are stacked, one above another, inside tall metalracks like high-rise condos. When you lean close and peerinside, you’re likely to get a visual jolt.

The cold, hard sterility of biomedical science is, you realize,wrapped around hundreds of breathing beings with whiskeredsnouts and beating hearts. They cuddle together for warmth andcompanionship. They look out at you with the pinkish eyescharacteristic of albino Strain F344, understanding nothingabout the scientific enterprise in which they play the leading role.

A Fish Like MeSo why do scientists work with animals? What can rats (Rattusnorvegicus) or zebrafish (Danio rerio), seemingly so far fromHomo sapiens on the tree of life, reveal about human health anddisease? Turns out, many basic biological processes such as celldivision, organ differentiation, gene mutation and diseaseformation play out similarly across species. That’s why a rat or amouse or a fish can act as a stand-in for a human in studies onmicronutrients, obesity, aging, ALS, cancer, drug efficacy,infectious disease and any number of other biomedical questionsunder investigation at Oregon State.

A Whole Lot of SeriousnessWhen researchers use rats, mice or other species to studyprocesses that mimic or parallel human biology, they call it a“model.” One common model is a “knockout mouse.” It workslike this: To gauge how certain genes affect certain bodilyfunctions or disease processes, researchers “knock out” orsilence the targeted gene and then study what happens when themice get, for instance, a high-fat diet or a hormonal supplement.Knockout mice are used at OSU to study bone growth, aging,obesity, immunodeficiency and many other intricate areas ofhuman health.

But complex animals like mice and rats are used only whenthere’s no other way to investigate the question at hand, Durkeestresses. Indeed, basic biomedical research begins with cells in atest tube. Only after experiments have shown great promise doscientists advance to animal work. And then, only after the

animal studies achieve high rates of treatment success or cures— along with low risks for harm — do scientists go on toconduct experiments on humans. Steve Durkee’s mother was asubject in one of those experiments, which researchers call“human” or “clinical” trials, when she was battling breast cancer.

Durkee likes to direct people to the AAALAC website’s long list ofNobel Prizes in medicine and physiology over the past 110 years.Without the use of lab animals, Frederick Banting and JohnMcLeod wouldn’t have discovered insulin and the mechanism fordiabetes, winning the Nobel in 1923. Alexander Fleming, ErnstChain and Howard Florey wouldn’t have discovered penicillinand its curative powers. Typhoid and yellow fever would still beraging across the land.

But Banting and McLeod’s methods with dogs, rabbits and fishprobably would fail to pass muster with today’s regulatingagencies. It’s not only federal regs that have changed — it’s themoral, philosophical and ethical sensibilities of Americans towardcreatures of all kinds. Oregon State biomedical ethicist CourtneyCampbell has seen a sea change over the past decade and a half.

“Nothing is more importantin an animal study than the animal itself,” says Steve Durkee.

“There’s a generational change going on,” says Campbell, whohelped lead a series of national ethics workshops for land grantfaculty in the 1990s. “The change isn’t limited to animal researchat universities — it’s also about food and entertainment andsports. It’s about the treatment of animals at zoos, circuses,aquariums, rodeos. “It’s about our diets — how veganism andvegetarianism were way out in the ‘fringy granola movement’ notthat long ago. “We haven’t done a complete cultural 180, butthere is definitely a new moral consciousness.”

At the end of the dayIn the rat room, the “Bruss” eaters live alongside the “brocc”eaters (broccoli sprouts) and the “fat” eaters (high lipids). There’sa control group, too, which eats regular rat chow. That’s soDashwood can compare the health impacts of an ordinary dietagainst those of the special diets. At the study’s start, all theanimals were injected with the carcinogen found in charred meat— a known cancer-causing compound to which most Americanshave been exposed in barbequed burgers or grilled steaks. Oncethe study is over, the animals will be euthanized, humanely, instrict accordance with the protocols set out by the AmericanVeterinary Medical Association. The researchers will thencompare the number and size of colon tumors among the fourgroups to find out whether eating sprouts made a difference.

When they talk about ending the lives of animals used inbiomedical research, Diggs, Durkee and Murray all express aresigned sadness. None of them could do their jobs without atotal conviction that scientific discovery justifies the animals’demise — that the death of a rat may someday save the life of achild. Still, it’s unsettling. “Nobody likes it,” muses Murray, hisattempt at matter-of-factness not 100 percent convincing. “But itis what it is.”


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A Kansas State University professor’s research improving post-surgery pain treatment and osteoarthritis therapy in dogs may helpdevelop better ways to treat humans for various medical conditions.

From the use of hot and cold packs to new forms of narcotics,James Roush, professor of clinical sciences, is studying ways tolessen pain after surgery and improve care for small animals,particularly dogs. He is working with the clinical patients who cometo the College of Veterinary Medicine’s Veterinary Health Center.

Because humans and dogs experience some diseases in similarways, his research may improve how doctors and physiciansunderstand human health, too.

“Several of our projects have human applications, particularly oneinvolving intra-articular prolotherapy,” Roush said.

Here’s a closer look at three of Roush’s current projects:

A recent project with Ralph Millard, former Veterinary Health Centerresident, focuses on ways that hot packing and cold packing affect tissuetemperature in beagles and beagle-sized dogs after surgery.

After surgery in both humans and dogs it is common to put a coldpack or hot pack on tissue to prevent and reduce swelling. Howlong the pack is used and what type of cold or hot pack is useddepends on the type of injury and surgery. Roush said that nostudies have looked at how deep in the tissue the packs affecttemperature and how long the packs must be applied so that thetissue reaches a desired temperature.

The researchers studied the temperature and tissue depth that hot andcold packing affected and the time it took to reach that temperature.

“We found that you don’t really need to cold pack anything longerthan 10 minutes because there is not a great change intemperature after that,” Roush said.

When tissue is cold packed, it will stay cold for a while after the icepack is removed. But when tissue is hot packed and the pack isremoved, the tissue temperature will return to normal much morequickly. Leaving the hot or cold pack on the tissue longer than 10minutes will extend the time that the tissue stays at the same hot or

cold temperature, Roush said. There just will not be a great changein temperature after 10 minutes.

The same technique of hot and cold packing after surgery is alsoused in humans. Although more research in humans is needed,Roush said there is a strong possibility that a similar 10-minute timeframe for hot and cold packing may apply to humans as well.

The research appears in two upcoming publications in the Journalof Veterinary Research.

For another project, Roush and Matt Sherwood, Veterinary Health Centerresident, are using a mat system to study lameness and osteoarthritis indogs. When dogs step on the mat, it measures the pressure in their step.

The mat system is a useful clinical tool for evaluating anddeveloping treatment of lameness, Roush said. Roush andSherwood are using the mat for measuring lameness anddetermining in which leg the lameness is worse.

“We’ve designed the study to help improve osteoarthritistreatment,” Roush said. “We will also use it to measure clinicalpatients when they come in for regular checkups. We can measuretheir recovery and a variety of other aspects: how they respond tononsteroidal anti-inflammatories, how they respond to narcotics orhow they respond to a surgical procedure that is designed to takethat pressure off the joint.”

Roush also is working with Marian Benitez, Veterinary Health Centerresident, on an analgesic pharmacology study. Rose McMurphy, professor ofclinical sciences, and Butch KuKanich, associate professor of anatomy andphysiology, are also involved.

The researchers are studying the effectiveness of a painkiller used totreat dogs and researching potential alternatives to the drug. Thesame drug also is commonly used to treat pain in humans.

“To achieve the drug’s effect, the dosage in dogs is much higherthan in people,” Roush said. “It also may not be a very goodanalgesic in dogs. We want to see if there is an alternative thatrequires smaller doses and does not have not as much of adiscrepancy for patients.”

A healthy bond:By improving paintreatment, therapy in dogs, researchoffers medicalinsight for humansby James Roush

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THE SOFT BUZZING OF MOSQUITOESFILLS THE AIR while spotted moths with 4inchwingspans flutter past. Bluegreen caterpillarsas thick as fingers crawl in a living carpet.

Summer picnic nightmare? Nope—thismotley collection of insects lives inside plasticcages in Estela Arrese’s biochemistry lab atOklahoma State University (OSU).

Arrese studies these and other insects to learnhow they—and we—store food as fat andlater break it down for energy. Her discoveriescould lead to new ways for farmers to protecttheir crops from pests, and for health officialsto combat mosquito-borne diseases likemalaria and West Nile virus.

Not only that, but studying these little critterscould one day improve our understanding ofdisorders like diabetes, obesity and heartdisease, which relate to how we store and use fat.

Saying that we can learn about humanbiology from mosquitoes and moths“sounds kind of crazy,” Arrese readilyadmits.

But the argument is hardly outrageous.Scientists have been studying insects tobetter understand human biology for morethan 100 years (see “Fruitful Work,” page6). While we may not look alike,caterpillars, flies and humans use similarmethods to regulate fat at the molecularlevel.

“It’s very exciting,” Arrese says of her work.“We are learning new things all the time.It’s a good time to be in this field.”

Fats Are UsAll creatures—people, insects, even plants—need fat to survive.

Most of the fats we eat are calledtriglycerides. Triglycerides give us more thantwice as much energy as carbohydrates orproteins. But before we can use that energy,our bodies must break down the fats.

When we digest triglycerides, they get splitinto their component parts: three fatty acidsand a carbohydrate. This splitting is calledlipolysis (lipfor “lipid,” or fat, and lysis for“split”), and the enzymes that do the splittingare called lipases.


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Once they’ve been absorbed into ourintestines, the fatty acids are recombinedinto triglycerides and shipped out to ourcells through the bloodstream. Some ofthe fat gets used for energy right away.The rest is stored inside cells in blobscalled lipid droplets.

Until the 1990s, researchers thought lipiddroplets were just beads of oil passivelyfloating around in cells. Then theydiscovered that the droplets are actuallydynamic structures that help regulatewhen stored fat gets broken down forenergy. Now, lipid droplets have takencenter stage in fat research.

When the body needs extra energy —forinstance, when we run a marathon—hormones direct lipases to break down thefats stored in lipid droplets and wash themback into the bloodstream.

The cycle of making, breaking, storingand mobilizing fats is at the core of howhumans—and all animals— regulate theirenergy. An imbalance in any step canresult in disease, including obesity anddiabetes. Having too many triglycerides inour bloodstream raises our risk of cloggedarteries, leading to heart attack andstroke.

Despite their importance, no one yetunderstands exactly how fat storage andmobilization work.

For instance, insects like silkworms andosquitoes store up lots of fat when they’reyoung by eating nectar or leaves. Theyuse that fat later when they metamorphoseinto their adult forms and start flying.

They also burn fat when they lay theireggs. If researchers could block themovement of stored fat to the insects’ovaries, they could interfere withegglaying and stop the bugs fromreproducing. That could have an immenseimpact on pest and disease control.

Protein Puzzle PiecesBut before any of that can happen, saysArrese, “We need to study a lot and haveinformation at the molecular level.”

It’s like an enormous, living puzzle, andArrese is trying to identify a few proteinpieces and figure out how they fit together.

She has already discovered the function of a protein called Lsd1 found in lipiddroplets.

Arrese also works with the main lipaseinvolved in fat regulation in insects.Fittingly, it’s called triglyceride lipase, or TGL. Now she’s looking into how TGL works.

She has a lot of questions: Does TGLactivate fat mobilization, block it or helpanother protein? Do other proteins helpregulate TGL? What is the function ofanother lipase, called ATGL? How doesthe Lsd1 protein help control lipolysis, andis it a critical target for disease control?What does its sister protein, Lsd2, do?

“In a cell, there are so many proteins, it’shard to tell which protein really does thework,” says physicist Donghua Zhou, oneof Arrese’s colleagues and collaboratorsat OSU.

These lipid droplets store fat in the cells ofthe tobacco hornworm, Manduca sexta.

To determine which proteins perform whatjobs, biochemists like Arrese have toisolate and purify each one in a test tubebefore conducting extensive experiments.

And purifying proteins is not easy, explainsJosé Luis Soulages, who collaborates withArrese in the university’s biochemistrydepartment—and who also happens to beher husband.

“After a lifetime, you may say that you’vefound four key proteins,” he says. “Andthere are probably 100 more. But theirfunctions would never be known withoutthe discovery of those first four.” ➤

Fats and Flies BY STEPHANIE DUTCHEN

Arrese studies insects, including the tobaccohornworm shown here (counterclockwise from top)as a c aterpillar, pupa and adult moth.

These lipid droplets store fat in the cells of the tobacco hornworm, Manduca sexta.

– Photo by ESTELA ARRESE


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‘Shockingly Good’It’s especially tough to purify Arrese’sproteins because they don’t survive longoutside cells and because they’re foundin oils. Most biochemistry tests aredesigned for waterbased molecules—andas you know if you’ve ever shaken abottle of salad dressing, oil and waterdon’t get along.

That doesn’t deter Arrese. Her lab wasthe first to purify TGL from fat tissue ininsects as well as Lsd1 and Lsd2, and shecontinues to purify proteins with acharacteristic drive for perfection.

“When she brings me molecules, they are impeccable,” says Steve Hartson, a biochemist at OSU who collaborateswith Arrese.

An advantage to using insects like fliesand caterpillars is that Arrese can raisehundreds of them in the lab and collectplenty of the proteins she wants—manymore than she could from rats or people.

“It puts her on the edge of what’spossible,” explains Hartson.

Arrese aims to learn more about theproteins by examining their structuresatom by atom.

But she can’t use one of the mostcommon protein imaging techniques,Xray crystallography, because herproteins don’t naturally form the neededcrystal structures. Nor can she use theother common technique, called solutionnuclear magnetic resonance (NMR),because the proteins are too big.

So Arrese collaborates with Zhou, whouses related technology called solidstateNMR. She hopes that soon, she willfinally be able to see the structures of herproteins in 3D.

It’s definitely not a job for people withshort attention spans.

Luckily, says Soulages, his wife iswellsuited to the work—she hasextraordinary concentration, keen powersof observation and a talent for planningefficient experiments. The combination oftalent and commitment allows her topursue a single scientific question fordecades without losing focus.

“I think very few people can do that,” hesays. “In science, sometimes we aretempted to switch subjects or even fieldsbecause things get hard. She just puts inmore hours and tries alternateapproaches.”

And it pays off. Hartson says, “She’s ashockingly good biochemist. She doessome incredible things with molecules.”

Hartson recalls being “a little intimidatedby her at first. She was very focused ondoing science. But then she makes adiscovery or [gets] key data and breaksinto this radiant, beaming person. She’sa real pleasure to work with.”

Getting Her Hands DirtyArrese acquired her stringent work ethicas a young girl, when she spent summerson her family’s farm in rural Pehuajo,Argentina.

What do I need to study to look throughmicroscopes a lot?

“My father always wanted us to be busydoing useful things,” she says.

For example, she and her brother werecharged with taking care of a bigvegetable garden.

Rather than resenting the work, Arresesays, “I realized I really liked it. I needthat. I need to get my hands dirty.”

It’s still the case metaphorically in thelab, where she is always ready with a pairof rubber gloves if she needs to dive intothe insect cages or help her students withan experiment.

It’s also true at home, where shecultivates a thriving flower garden despiteOklahoma’s unforgiving climate—drysummers, harsh winters, tornadoes andspring hail that drives Arrese outside tothrow a protective tarp over her tulipsand roses.

“Whatever it takes to have flowers, I willdo it,” says Arrese. “I don’t care if it’s achallenge. Because I was raised on thatfarm, I am used to a lot of work.”

Dedication to ScienceArrese’s father, a veterinarian, also raisedher to love science.

She always felt comfortable in the lab heset up in their house, wandering amidstthe pipettes and tubes, the microscopeand canisters of liquid nitrogen. By theage of 10, she was helping her fatherand pretending to be a scientist herself.

Her favorite task was looking through themicroscope. When she was in middleschool, she asked her father, “What do Ineed to study to look throughmicroscopes a lot?”

His answer: biochemistry.

“And that,” she says, “is what I ended up doing.”

The path wasn’t an easy one.

She gave birth to her first child while she was still working on her Ph.D.thesis—just before her husband moved to the United States.

When she and her daughter joined herhusband in Tucson, Arizona, she leftbehind her extended family, her languageand her culture.

In Argentina, she had taught herself toread and write English with a dictionary inone hand and scientific journal articles inthe other. But she arrived in the U.S.without knowing how to speak thelanguage. She still struggles to speak asfluently—and as much—in English as shedoes in her native Spanish.

It’s like an enormous, living puzzle.

Arrese as a child.

– Photo by AMERICO ARRESE


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Despite the communication barrier, Arrese loves English. “It’s moreprecise,” she says. “I am 50—maybe when I retire, I will study it.”

Experimenting in the KitchenAlong with the lab, Arrese has always felt comfortable—andenjoys experimenting—in the kitchen.

When she has to follow a recipe, she says, “It’s a disaster. I want tochange it. I want to know what will happen if I add this or that.”

Her husband agrees: “She has to see how people are cooking,and then she works it out on the bench—on the stove.”

Some of her favorite dishes are cod and paella. Among herappreciative consumers are her husband, her two daughters andher students.

“I love cooking because I like good food, because I want to feedmy family and also because I need that—to go and createsomething,” says Arrese.

Growing up, she learned how to prepare not only an eclecticarray of Argentine dishes, but also Spanish food from an aunt whowas a chef and Italian meals from her immigrant grandmother.

When she relocated to Tucson, she promptly pestered a neighborto teach her Mexican cuisine.

When the family moved again, this time to Oklahoma, a handfulof graduate students from India joined her lab and helped herfulfill a decadesold desire to learn to cook Indian food. Wheneverthese students traveled home, they would bring back suitcasespacked with spices for her.

Now Arrese has three graduate students from China, and she’slooking forward to once again expanding her culinary horizons.

Extended FamilyWhen she isn’t grilling her students for cooking tips, Arrese caresfor them as though they were her extended family.

“She nurtures her students…in such a way that the excitement ofscience never wears off and critical thinking is developed,” saysAlisha Howard, who did undergraduate research in Arrese’s labbefore earning her Ph.D. with Soulages.

“She seems to be very invested in our future as scientists and asadults,” agrees Zach Hager, a college senior in the lab.

Hager particularly appreciates Arrese’s eagerness to workalongside her students at the bench.

Colleague Steve Hartson says that Arrese’s “tremendousoneonone mentoring and intellectual and moral support” are aninspiration to her students and ensure that they achieve theirpotential. “They’re excited by the science they’re doing, and theygo on to very successful careers,” he reports.

Her students also admire the way shebalances the jobs of lab head and mother(her daughters are now 13 and 20).

It helps to have her husband nearby. Sheand Soulages often take turns running longexperiments or helping their kids with dailyroutines.

Her ultimate goals, she says, are to raise her children to be healthy and have goodlives, and to find out what her handful ofproteins do.

Soulages describes his wife’s philosophy notas seeking an earthshaking discovery, butas putting in her grains of sand, knowingthey will last a long time. He could be talking aseasily about her children and students asabout her science.

For her part, Arrese gives the same adviceto all:

“Identify [your] passion and follow it. Whenyou really like what you do, it becomeseasy. Live and work with passion andquality.”

Identify [your] passion and follow it.

Arrese (right, in blue) teaches her studentsto be efficient and excited about science.

– Photo by ESTELA ARRESE


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Tackling diabetesTEAM HOPES RESEARCH FROM ANIMALS CAN

LEAD TO HELP FOR HUMANS

Véronique Lacombe, DVM, Ph.D., brought a long history ofcomparative medicine research with her when she joined OSU’sveterinary center as an associate professor in physiologicalsciences. Since she was a veterinary student, Lacombe has had aninterest in metabolism, the complex process the body uses to turnfood into energy. Specifically, her fields of interest include skeletaland cardiac muscle energetics, glucose transport during diabetes,insulin resistance using small- and large-animal models, as well ascardiovascular complications during diabetes.

The main mission of Lacombe and her team is researchingmechanisms underlying diabetes, a complex disease for which thereis no cure.

“Diabetes is an epidemic disease that affects more than 250 millionpeople with almost 10 percent of the population affected in theU.S., and it is expected that the worldwide prevalence will rise to450 million by 2030,” she says. “As a result, the disease imposes aconsiderable medical and economic burden on societies. My lab isinvestigating the regulation of glucose transport in insulin-sensitivetissue. In other words, we are looking at how the glucose (i.e.,sugar) in the bloodstream transfers to tissue. This process is themetabolic bottleneck for glucose utilization and fuel production. Inaddition, this process is altered in people who have diabetesbecause they have improper production and/or action of insulin, ahormone that is necessary to make that transfer.”

While there is no drug to cure diabetes, human diabetes can beregulated and monitored to avoid complications.

“With a diet regimen, exercise and weight loss, diabetics can helpmanage their disease. When skeletal muscles contract, that processhelps transport glucose into cells. Exercise can speed glucoseuptake in muscles. However, the process by which contractionenhances glucose transport is unknown, and it is one of theresearch focuses of my laboratory. Findings from this research couldlead to the discovery of a cure for diabetic patients.”

Lacombe says glucose is one of the main sources of fuel for thebody, and the uptake of glucose from the blood into the cell and itsutilization by the cell to produce energy is similar across all species— human and animal.

“Because the process is similar, we use both small- and large-animal models in my lab, spanning from mice to horses. If we findmechanisms responsible for diabetes using these species, it couldalso have an impact on human health, a concept referred to as onehealth, one medicine,” she says. “For example, in mice, we canupregulate a protein potentially implicated in the transport ofglucose to see if it will help prevent diabetes. As a result, we havenow established a line of mice that are resistant to diabetes.

DR. MELODY DE LAAT (LEFT)DEMONSTRATES THE NEW

MICRO-ULTRASOUND MACHINEWITH DR. VERONIQUE LACOMBE(CENTER) AND BRITTANY EVANS.

– Photo by PHIL SHOCKLEY / UNIVERSITY MARKETING


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Big pictures from little things BY MATT ELLIOTT

NEW MICRO-ULTRASOUND MACHINE GETS CLEAR PICTURES FROM INSIDE SMALL CRITTERS

The Center for Veterinary Health Sciences purchased a state-of-the-art ultrasound machine in2011 that greatly expands researchers’ ability to image small animals.

The machine, the VisualSonics Vevo 2100, allows vet school scientists such as its operator, Dr.Véronique Lacombe, to see into the organs and systems of lab animals as small as mice andrabbits. The high-frequency digital imager adds to the vet school’s growing list of imagingtechnology that already includes MRI and CT scan machines, and a human-size ultrasoundmachine available at the teaching hospital.

Ultrasound uses sound waves to create images of how things work inside bodies by translatinghow the waves pass through different objects into an image. They are used in everything fromimaging fetuses and the hearts of people suffering from cardiovascular disease to cancer.

It’s easy to envision their usefulness in veterinary medicine as well, including the booming realm ofcomparative medicine and preclinical research that focuses on problems afflicting both animalsand humans. Naturally, it makes sense to use animals to model human systems in research.

The problem is that large ultrasound machines operate at a lower sound frequency to passthrough larger patients, such as a human or a horse. It is not as helpful to image a mouse or a ratbecause the image quality wouldn’t be as good.

“With this micro-ultrasound machine, you can’t go very deep, but you’re going to have aspectacular resolution of the specific area that you’re examining,” Lacombe says. “The advantageof this machine is that it has the same features as a human ultrasound machine, but it has beendesigned to image all the organs of small animals, including of early embryonic and neonatalmice.”

The Vevo 2100, believed to be the only one in Oklahoma, is in Lacombe’s ComparativeMetabolism Laboratory in the vet school’s physiology department.

It has already proven helpful in her research.

As diabetes is one of Lacombe’s chief areas of interest, she often works with small animals, suchas mice and rats as models of human metabolic diseases.

One of the increasingly common condition’s accompanying diabetes is heart disease. Previously,she had to use human ultrasound machines in her animal models that were not as useful in herwork.

This machine lets her look at everything from the shape of the heart to how effectively it pumpsblood, as well as how the heart tissue contracts and relaxes during each heartbeat, letting her“detect subtle heart dysfunction very early on in the process of the disease in our diabetic mice,”Lacombe says.

“Also, with this high resolution, you can also inject anything you want into a targeted site, includingthe brain and the spinal cord. You can also use micro bubble technology,” she says. “You cancreate small bubbles of air that will go through the heart or blood vessels to mark the passage ofwhat you’re trying to track.”

She can also look at blood flow into organs to track vascular diseases. It also lets her deliver geneand stem cells at a targeted site and measure things such as gene therapy effectiveness. And 3-Dreconstructions of organs are possible.

“It’s a huge improvement since this micro-ultrasound machine greatly expands our understandingof the physiologic and pathophysiological processes in small animal models.” Lacombe says.

The Vevo 2100 is available for any researcher in the veterinary college or on OSU’s campus touse, she says.

“It can be used by any investigators on campus since it is a common equipment. I think the morepeople we have using it from different areas of research, the better it would be. I think it couldeven generate some collaborations between the veterinary school and other colleges on campus.”

Like people, horses can becomeobese, which can lead to ametabolic disorder such as insulinresistance. Interestingly, cats can gettransient diabetes, where thediabetes goes into spontaneousremission. Unlike the rodent modelsused to study diabetes, which aregenerated from inbred strains oflaboratory mice, these naturallyoccurring models are geneticallydiverse and exposed to many of thesame environmental factors thathumans are, and they are greatmodels of metabolic disorders.

By transcending species boundariesto include the study of spontaneousand experimental models of humandisease, research in comparativemedicine can lead to excitingdiscoveries that will benefit bothpeople and animals.”

Lacombe firmly believes thatveterinarians’ thorough training puts them in a unique position toimprove research and help society.

“Veterinarians have such a broadtraining. We have to know all thedifferent species from fish toelephants. That vast knowledge canbe applied in comparative medicineresearch. It is a career path withmany rewards that many veterinarystudents don’t really think about.”

After earning her DVM degree,Lacombe completed her residency in large-animal internal medicineand worked as an equine clinicianbefore focusing on research.

“As a clinician first and a scientistsecond, I am a better researcher and ask questions that are clinicallyrelevant. Clinical veterinary medicineand research are similar processes.In both cases you start with aproblem. In clinical veterinarymedicine, you have a list of differentdiagnoses that could be causing the problem. In research, you havedifferent hypotheses you want toprove. One by one, you check themoff the list in both areas. In clinicalveterinary medicine you have a final diagnosis, and you treat it. In research, you have the correcthypothesis, and you find a cure oryou take one step closer to finding acure. We are responsible to nourishthat aspect of the veterinaryprofession and train the nextgeneration of veterinary scientists.”


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Johns Hopkins scientists report what is believed to be the first evidence that complex, reversible behavioralpatterns in bees – and presumably other animals – are linked to reversible chemical tags on genes.

The scientists say what is most significant about the newstudy, described online September 16 in NatureNeuroscience, is that for the first time DNA methylation“tagging” has been linked to something at the behaviorallevel of a whole organism. On top of that, they say, thebehavior in question, and its corresponding molecularchanges, are reversible, which has important implications for human health.

According to Andy Feinberg, M.D., M.P.H., Gilman scholar,professor of molecular medicine and director of the Centerfor Epigenetics at Hopkins’ Institute for Basic BiomedicalSciences, the addition of DNA methylation to genes has long been shown to play an important role in regulatinggene activity in changing biological systems, like fatedetermination in stem cells or the creation of cancer cells.Curious about how epigenetics might contribute to behavior,he and his team studied a tried-and-true model of animalbehavior: bees.

Working with bee expert Gro Amdam, Ph.D., associateprofessor of life sciences at Arizona State University and theNorwegian University of Life Sciences, Feinberg’s epigeneticsteam found significant differences in DNA methylationpatterns in bees that have identical genetic sequences butvastly different behavioral patterns.

Employing a method that allows the researchers to analyzethe whole genome at once, dubbed CHARM (comprehensivehigh-throughput arrays for relative methylation), the teamanalyzed the location of DNA methylations in the brains ofworker bees of two different “professions.” All worker beesare female and, within a given hive, are all geneticallyidentical sisters. However, they don’t all do the same thing;some nurse and some forage.

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Bees DECIDE WHAT to BeBy Vanessa McMains, Ph.D.

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Nurses are generally younger and remain in the hive to take care of the queen and her larvae. When nurses mature, they becomeforagers that leave the hive to gather pollen and other supplies for the hive. “Genes themselves weren’t going to tell us what isresponsible for the two types of behavior,” Feinberg says. “Butepigenetics – and how it controls genes – could.”

Feinberg and Amdam started their experiment with new hivespopulated by bees of the same age. That removed the possibilitythat any differences they might find could be attributed todifferences of age. “When young, age-matched bees enter a newhive, they divvy up their tasks so that the right proportion becomesnurses and foragers,” explains Amdam. It is these two populationsthat were tested after painstakingly characterizing and markingeach bee with its “professional,” or behavioral, category.

Analyzing the patterns of DNA methylation in the brains of 21nurses and 21 foragers, the team found 155 regions of DNA thathad different tag patterns in the two types of bees. The genesassociated with the methylation differences were mostly regulatorygenes known to affect the status of other genes. “Gene sequenceswithout these tags are like roads without stop lights – gridlock,”says Feinberg.

Once they knew differences existed, they could take the next step todetermine if they were permanent. “When there are too few nurses,the foragers can step in and take their places, reverting to theirformer practices,” says Amdam. The researchers used this strategyto see whether foraging bees would maintain their foraging genetictags when forced to start acting like nurses again. So they removedall of the nurses from their hives and waited several weeks for thehive to restore balance.

That done, the team again looked for differences in DNAmethylation patterns, this time between foragers that remainedforagers and those that became nurses. One hundred and sevenDNA regions showed different tags between the foragers and thereverted nurses, suggesting that the epigenetic marks were notpermanent but reversible and connected to the bees’ behavior andthe facts of life in the hive.

Dramatically, Feinbergnoted, more than halfof those regions hadalready beenidentified among the155 regions thatchange when nursesmature into foragers.These 57 regions arelikely at the heart ofthe different behaviorsexhibited by nursesand foragers, saysAmdam. “It’s like oneof those pictures thatportray two different images depending on your angle of view,” she says. “The bee genome contains images of both nurses andforagers. The tags on the DNA give the brain its coordinates so that it knows what kind of behavior to project.”

The researchers say they hope their results may begin to shed lighton complex behavioral issues in humans, such as learning, memory,stress response and mood disorders, which all involve interactionsbetween genetic and epigenetic components similar to those in thestudy. A person’s underlying genetic sequence is acted upon byepigenetic tags, which may be affected by external cues to changein ways that create stable – but reversible – behavioral patterns.

Authors on the paper include Brian Herb, Kasper Hansen, Martin Aryee,Ben Langmead, Rafael Irizarry and Andrew Feinberg from The JohnsHopkins University, and Florian Wolschin and Gro Amdam of theNorwegian University of Life Sciences and Arizona State University.

This work was funded through the NIH Director’s Pioneer Award through the National Institute of Environmental Health Sciences(#DP1ES022579), the Research Council of Norway and the PewCharitable Trust.

– Photos by Christofer Bang

to feed and take care of the queen and her larvae. Forager bees are responsible for gathering pollen and nectar.

The researchers say they hopetheir results may begin to shedlight on complex behavioral issuesin humans, such as learning,memory, stress response andmood disorders, which all involveinteractions between genetic and

epigenetic components similarto those in the study.


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Helen was a typical mom of the 50s and 60s with four kids who all had busylives. By 1974, one son was married with two youngsters of his own. Onedaughter was just out of college and the two youngest children were in college.One fall day that year, they each received a call that their mother had beendiagnosed with a brain tumor. Seven hours of surgery, months of radiationtherapy, and nine months of worry ended in Helen’s death at the age of 55.

Immediately after the operation, the surgeon had said that the kind of tumorthey found was like “spilled milk.” Today, that’s still most often the story of whathappens to people with the deadliest form of brain tumor, glioblastoma, alsoknown as a stage IV glioma or stage IV astrocytoma. Sen. Ted Kennedy lived for14-1/2 months with one, dying in 2009 even after receiving top-of-the-linemedical care.

A team of scientists and surgeons at Virginia Tech-Wake Forest University Schoolof Biomedical Engineering and Sciences have teamed up to use a newtreatment technique on dogs, with the expectation that within a few years theymay be performing clinical trials on humans with glial cell tumors.

Glioblastomas in canine patients are almost identical to those found in humansand, in fact, occur at least three times more often in dogs. And sadly, mortalitypatterns for dogs also are almost identical when corrected for normal longevity;dogs survive a few weeks to a few months and humans about nine months withsurgery, chemotherapy, and radiation, and 15 months when a drug calledtemozolomide is added. It’s these fatal facts that give researchers hope that theycan learn from our four-legged pals how to beat this deadly demon for bothpeople and pups.

“Dogs are the holy grail of a spontaneous model for glioblastomas because thetumors develop just like in people,” says John Rossmeisl, the neurosurgeon inthe Virginia-Maryland Regional College of Veterinary Medicine who is using thenew procedure on dogs suffering from brain tumors and some other forms ofcancer. “We don’t know why these tumors develop. Some of it may bechemicals, cleaning products. Dogs are so similar in makeup to humans thatthey can be the environmental sentinels for people.”

The brain is a mysterious place with an estimated 100 billion neurons, or nervecells, that fire up to make your thoughts swirl, your senses work, and your bodymove. Researchers don’t know exactly how the brain functions and they don’tknow why glioblastoma multiforme happens. But when the type of glial cellscalled astrocytes start multiplying uncontrollably inside a person or dog’s skull,it’s difficult to stop. One reason is that these types of tumors weave theirtentacles into the surrounding tissue. Traditional radiation and chemotherapycan’t pinpoint tumor cells precisely enough to prevent damage to other tissues.

Trying something newA new treatment, non-thermal irreversibleelectroporation (N-TIRE), uses two electrodesabout one millimeter in size, approximatelythe same as a needle used for thick thread,placed directly on the tumor cells. Very shortpulses of electricity shoot through the deviceinto the cancer cells. The electric pulse is soshort that it raises the temperature of thewire by only about one-quarter of a degree.

One of the first patients treated with N-TIREby the Neurology and Neurosurgery Serviceat the veterinary college in Blacksburg, Va.,was a 12-year-old dog sent there becausehe was having vision problems, seizures,behavioral changes, and was unsteady onhis legs.

These are some of the same symptoms thathumans often exhibit. The tumor takes upspace in the inelastic skull that the brainneeds to function. So the brain is crowdedand some of its normal work is blocked,preventing transmission of signals thatcontrol everything from your eyes to yourfeet. In Helen’s case, she was having somevision, odor perception, and memoryproblems before the operation, and whenthe cancer cells not excised during surgeryor killed by radiation grew into a tumoragain, she lost her ability to speak, againhad vision problems, and was unsteady onher feet until finally bedridden.

Rossmeisl’s canine patient was diagnosedwith a glioblastoma that was too large andrapidly growing to allow for standardsurgery. This made the dog a candidate forN-TIRE.

Two days after the procedure, a magneticresonance image (MRI) showed that thedog’s tumor had shrunk to 75 percent itsoriginal size. He was put on a regimen of

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radiation, and by four months post-procedure, the family petwas in complete remission.

Non-thermal irreversible electroporation actually makes holesin the cancer cells that cause them to die. But because thistreatment does not generate heat and because it can hone inon the tumor so accurately, it doesn’t damage the normal cells,nerves, or normal vascular system around the tumor.

“What we’re really doing is changing the cancer tissue’sproperties,” says Rafael Davalos, co-inventor of the procedure.“Whatever area is treated, we can pinpoint it to half amillimeter. Also, it is super-fast, so there’s no heating and no scarring.”

The surgeon can see the placement and progress of theelectrodes in real time by using an ultrasound or MRI, saysDavalos, a biomedical engineer in the Virginia Tech-WakeForest University School of Biomedical Engineering andSciences, who attends every procedure that Rossmeisl doesusing N-TIRE.

Currently, the standard follow-up protocol of radiation andchemotherapy is used on the canine patients after having aprocedure using N-TIRE. Chemotherapy and radiation can be atwo-edged sword because they can damage normal tissue andcompromise the immune system. But the new treatment mayhave a two-fold benefit for the immune system.

“Irreversible electroporation seems to trigger an immuneresponse, so we may not need any drugs as part of thetreatment,” Davalos says. “That means your immune systemisn’t weakened any further.”

The veterinary college has successfully used N-TIRE on othertypes of cancers and other animals. Autumn, a chocolateLabrador retriever, was having trouble walking, so owner SherylCoutermarsh-Ott took her to the college for orthopedic surgery.They discovered that Autumn had an inoperable tumor on theinside of her thigh and groin.

“The tumor was wrapped around her femoral hip joint,” saysCoutermarsh-Ott. “The other options were not good. So, aftertalking to Dr. Rossmeisl, and reading some research papersabout N-TIRE, we went ahead. Now she’s doing absolutelyfantastic and is clear of cancer.”

Then there was the 11-year-old thoroughbred with anulcerated, growing cancer of the lip. The treatment with N-TIREalmost completely obliterated the tumor and the rest wasremoved with a carbon dioxide laser. The horse is now back tohorsing around.

The risksDespite those successes, the focus of the Virginia Tech-WakeForest team is on brain tumors, especially glioblastomas,because no really good treatment exists unless the tumors arediagnosed early, which happens rarely.

Like humans, dogs that develop glioblastomas tend to be older— seven years and up; for people it’s usually around age 65or older. More men than women develop this type of tumor.

According to the American Cancer Society, about 13,000Americans will die from brain tumors this year. More than 77percent of those tumors are glioblastomas, which have afatality rate of about 50 percent within 15 months and 75percent in 24 months. That’s actually a much better survivalrate than in 1974.

This type of brain cancer is called a primary tumor because itdevelops in the brain. Tumors that grow elsewhere in the bodyand spread to the brain are called metastatic tumors.

Because it’s often too late to save the life of someone with aglioblastoma, Rossmeisl is involved in a related project toinvestigate how to discover brain cancer early. Currently, by thetime symptoms appear, the tumor is often too large and hasinfiltrated too much of the brain to be treated effectively.

“Early detection is something that we really need,” Rossmeislsays. “We need something noninvasive that we can test for inblood or urine samples.”

The human sideDr. Thomas L. Ellis at Wake Forest is working with Rossmeisl,Davalos, and John Robertson to advance research on the useof N-TIRE. Robertson is a veterinary college professor ofpathology, the director of the Center for ComparativeOncology in the veterinary college at Virginia Tech, andcoordinator of the N-TIRE project. Ellis is a neurosurgeon andprofessor at the Wake Forest University Brain Tumor Center ofExcellence.

It’s Ellis who will take N-TIRE to clinical trials for people if givenapproval by the Wake Forest institutional review board and theFDA. If the procedure advances to human trials, it will first beused for patients who have recurrent glioblastomas. These arethe patients who have run out of treatment options.

Every month, a couple of patients come to Ellis with a diagnosisof glioblastomas. Although the cases are all different in someways, they’re also all the same in other ways.

“The causes for these tumors aren’t known; there are manyfactors but no known exposure pattern,” Ellis says. “Thesetumors are only rarely inherited. They occur sporadically inhumans and there are no known exposures to chemicals orother agents that predispose patients to develop them. Wedon’t know of any brain tumor that is connected with a priorhead injury.”

Scientists, including Ellis, don’t believe that primary braintumors are inherited; however, he has had two cases that seemto defy the odds. He had two siblings as patients who haddeveloped mirror-image glioblastomas, and a recent patient’sfather also had a glioblastoma.

In July 2009, a man from South Carolina who was havingtrouble with numbers and with spelling his sister’s name cameto Ellis. James Rollison thought he was having early onsetAlzheimer’s disease at the age of 58. He hadn’t suspected abrain tumor, although his father died of a glioblastoma. Butthat was what he had. ➤


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“The thought never crossed my mind that it was a GBM(glioblastoma multiforme) because my father’s doctor hadsaid there was no way it was hereditary,” Rollison says. Butwhen Rollison started having problems, his family doctorordered an MRI and then sent Rollison to a neurosurgeon.

Fortunately, Rollison’s tumor was one of the rare ones thatwas diagnosed early. The retired attorney credits his work andbeing attuned to his body to sending him to the doctor at thefirst indication something was wrong. Rollison doesn’t looklike, sound like, or act like someone who has had thedeadliest brain cancer possible.

Rollison was lucky because Ellis is a top-notch surgeon whogot the tumor relatively early and was able to remove 100percent of the visible mass. However, in order to treat themicroscopic tumor cells that inevitably remain after surgery,Rollison underwent the standard chemotherapy and radiationregimen. Now he goes back to Wake Forest every threemonths for a checkup with Dr. Glenn Lesser, of thehematology and oncology department of the university’sComprehensive Cancer Center, and has a follow-up MRI tomake sure the tumor hasn’t returned.

The past, present, and futureBut not all outcomes are like this and Ellis wants to have moreassurance that when he treats a brain tumor patient he’s givingthem a chance at life. “In the last 40 years, we’ve made littleprogress in treating these tumors,” he says. “Using radiationand chemotherapy the (survival) numbers are pretty dismal.”

In 1975, when Helen died, chemotherapy wasn’t commonbecause most drugs couldn’t pass through the blood-brainbarrier — a layer of closely spaced cells over the brain thatstops harmful substances from invading. According to theNational Cancer Institute, the rate of brain, spinal cord, andother nervous system cancers then was 5.9 per 100,000adults. Today the rate is 6.4 per 100,000.

Experts say the apparent increase in cases is mainly becauseof better diagnostic tools. Some of the treatments are alsobetter, such as the MRI that can be used both for diagnosticsand treatment. A functional MRI (fMRI) now exists that canmonitor neurological blood flow in real time. In addition,more drugs can cross the blood-brain barrier and somebetter ways to deliver drugs are under development. Still,improved technologies usually buy patients only a few monthsmore than they would have had 37 years ago.

“One of the more difficult parts of my job, that never gets anyeasier, is when a person comes in who was living a normallife the day before and I have to tell them that they have onlyone to two years to live,” says Ellis.

N-TIRE could give Ellis the advantage of reaching deep tumorsand giving uniform treatment, he says. So far the procedure ondogs has been “excellent. This is an amazing collaborationwith the vet school, Virginia Tech’s biomedical engineeringgroup, and Wake Forest. I truly believe that N-TIRE can becomean important new effective tool against glioblastomas.”

In the meantime, Rossmeisl and his team continue to use N-TIRE to treat dogs that have no other hope. “So far, theoutcomes with dogs have encompassed all the possibleclinical outcomes,” he says. He works with Robertson and

other collaborators to identify the reasons certain dogs, mostnotably dogs with snubbed muzzles and broad heads, such asboxers and Boston terriers, are more prone to primary braintumors. He also wants to make major strides in drug deliveryto overcome the blood-brain barrier so the medications godirectly where they are needed and stay there.

“We believe that glioblastomas have so many ways to evolveand to evade all the treatments we have now that if we canfigure out a way to beat those cancers we can cure anytumor,” Rossmeisl says.

Helen’s last Christmas was a joyful eventbecause, though she’d lost her hair fromradiation treatment and was wearing wigs and scarves, most of the family thought she was recovering – but, like the majority of thesetypes of tumors, it was discovered too late.

John H. Rossmeisl Jr., associate professor of neurology and neurosurgery in theveterinary college, does research onprimary brain neoplasms and traumaticbrain injury. He also sometimes works withthe college’s patients, such as Autumn,here with her owner, Sheryl Coutermarsh-

Ott. Rossmeisl treated Autumn for an otherwiseinoperable tumor in her groin using non-thermalirreversible electroporation. – Photo by Jim Stroup.

The electrode for irreversible electroporationfor treating cancer was developed by RafaelDavalos, assistant professor in the VirginiaTech-Wake Forest University School ofBiomedical Engineering and Sciences. – Photo by Jim Stroup.

Neurosurgeon Tom Ellis and his dog, Gandalf. – Photo by Jim Stroup.

Dr. Glenn Lesser of Wake ForestUniversity Medical Center exams JimRollison, who has been undergoingbrain tumor treatment. – Photo by John McCormick.

Robert E. Neal II, doctoral student in theVirginia Tech-Wake Forest UniversitySchool of Biomedical Engineering andSciences (SBES); John Robertson,director of the Center for ComparativeOncology in the Virginia-MarylandRegional College of Veterinary

Medicine; and Paulo Garcia, SBES postdoctoralassociate, position electrodes in a gel to look atcontact parameters as they test irreversibleelectroporation electrodes for potential applications. – Photo by Jim Stroup.


Tufts University

Sunrise sends streaks of red, violet andorange over the rippling surface of SanDiego Bay at Base Point Loma, where 25undergraduate interns are getting startedon the day’s work. They hose down agently swaying jigsaw of floating docksand pack 10-gallon thermoses withherring, capelin, smelt, mackerel andsquid that will fortify their charges—in thiscase, 60 bottlenose dolphins thatintermittently pop out of the water likeprairie dogs.

This seaside installation is home to theU.S. Navy’s Marine Mammal Program,which in the late 1950s studied howdolphins whip through water, with thegoal of improving torpedo, ship andsubmarine design. Today the Navy trainstwo species, the bottlenose dolphin andthe California sea lion, to help guardports, personnel and military vesselsaround the globe. They have served inVietnam and Iraq.

Because dolphins possess the mostsophisticated sonar known to man, theyare unrivaled in their ability to locate anddisable anti-ship mines and booby traps,even in the murkiest waters. Sea lions,with their superb low-light vision andsharp underwater directional hearing(something humans lack), can detect apotential enemy combatant swimmerapproaching a ship. Even moreimportantly, both species can makerepeated deep-water dives withoutsuffering the decompression sickness thathumans do.

What this means is that a single marinemammal, two handlers and a rubberboat can provide the same high level ofsecurity as a team of human divers andthe naval vessel, crew, physicians andmedical equipment needed to keep themfrom getting “the bends.”

So when it comes to providing for thesesoldiers of the sea, it’s nothing but top-flight.

The National Marine MammalFoundation is the nonprofit charitableorganization that provides medical carefor the Navy’s marine mammals. Cynthia

Smith, V99, is the executive director andmedical director of the foundation, whichcares for 120 Navy dolphins and sealions, ranging in age from newborns tosenior citizens. Foundation employees,the U.S. Navy, the U.S. Army, othercontractors and consultants staff the $20million operation at Point Loma, wherethe animals live and are trained in penssubmerged in San Diego Bay. Eleven full-time veterinarians and 160 trainers tendto every need of this marine securityforce. The dolphins and sea lions evenhave personal chefs: Army veterinariansinspect seafood from all over the world tosupply the one million pounds of fish theyconsume each year.

On this day, Smith, who has cared for theNavy mammals for more than a decade,arrives on base promptly at 7:30 a.m.Because the animals get so muchexercise swimming in the open ocean,her job is a lot like that of physician for ateam of Olympic athletes. It is alsosomewhat like treating patients fromanother planet; although scientists havebeen studying marine mammalphysiology for more than a century, littleis known about them relative to thelandlubber animal kingdom.

Take sea lions, which Smith says are likewater dogs, so “small animal medicinecomes in really handy.” Dolphins, on theother hand, she says, are more of aphysiological patchwork of people, pigsand cows.

It is those similarities between dolphinsand humans, learned by studying andcaring for these animals for nearly ahalfcentury, that have yielded anunanticipated benefit—a robust mountainof data that is helping to advance humanmedicine. One tantalizing outcomecurrently under investigation could leadto a treatment or cure for the 23.6million Americans who suffer from type 2 diabetes.

“Think about it,” says Stephanie Venn-Watson, V99, the veterinaryepidemiologist who heads the clinicalresearch enterprise at the NationalMarine Mammal Foundation.

“How many blood samples do we, aspeople, give in a lifetime? Well, comparethat with these animals, which routinelyprovide blood samples, have daily healthinspections, get annual ultrasoundexaminations and have access tonumerous diagnostic tests over 30 or 40years,” she says. “The power of thedatabase is endless. We haven’t found thelimit to the questions we are able to ask.”

The first fragments of that longitudinaldatabase were incubated at anamusement park in Santa Monica in1960. Sam Ridgway, an Air Forceveterinary officer stationed at nearbyOxnard, would tag along with the localvet, Robert M. Miller, to care for thedolphins and seals that performed atPacific Ocean Park. They published a fewof their cases in professional journals,including the first X-ray of a live dolphin.Ridgway, now regarded as the father ofmarine mammal medicine, was alsoassigned to serve on two naval bases,where he first met the military and civilianscientists who were interested in studyingthe mechanics of how dolphins swim sofast and dive so deep.

After completing active duty with the AirForce in October 1962, Ridgway washired as the animal health officer for theNavy’s first five bottlenose dolphins. Soonafter, he landed on a technique that isnow the cornerstone of the Navy’smarine mammal care regimen. In 1963,the Washington University physicist C.Scott Johnson trained a dolphin he wasusing in his auditory research so thatRidgway could do a complete physicalexamination without removing the animalfrom the water. Under the leadership ofRidgway, now president of the NationalMarine Mammal Foundation, the practiceof training marine mammals toparticipate in their medical care has beenrefined in the decades since.

“The goal is to keep the animalscomfortable and in their naturalenvironment at all times,” says CynthiaSmith. ➤

baywatchNavy dolphins are safeguarding our nationalsecurity — and advancing human medicine

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The Navy mammals can contract thesame illnesses that affect their relatives inthe wild: respiratory infections, diarrheaand other bacterial and viral diseases.The focus on preventive care and rapiddetection of illnesses has been sosuccessful, however, that the veterinariansare developing a geriatric medicineprogram. Many Navy dolphins are nowin their 30s and 40s; some are evenreaching 50. That’s ancient comparedwith dolphins in the wild, which typicallylive into their teens and 20s. Navy sealions live two or three times longer thantheir wild counterparts.

Dolphins give blood samples, have theirtemperature taken and provide fecal andurine samples—all while in the water. Assemi-land animals, sea lions undergo thesame tests while lounging on the f loatingdocks in San Diego Bay.

This morning, Smith pauses on the docknext to Mu, a 33-year-old dolphin. Murolls onto one side, bobbing in the wateras a veterinarian runs a portableultrasound over her belly. Four monthspregnant, Mu is one of six Navy dolphinsdue to calf this summer.

“It’s an exciting but anxious time,” saysSmith, who pauses to borrow a pair ofhightech sunglasses from the attendingvet, Forrest Gomez. The ultrasoundimage projects onto the inside of thelenses, allowing Smith to assess thegrowing fetus despite the harsh sunlightbouncing off the waves.

Prenatal ultrasounds are rare intraditional veterinary practice, so thefoundation team looks to humanmedicine as the standard. “We doultrasounds at least once a month at thisstage [of pregnancy],” says Smith. “Wecheck for a fetal heartbeat and look atfetal development. As the due dateapproaches, we do more frequentultrasounds.”

Pleased with Mu’s progress, Smith joinsthe rest of the medical team for adiscussion of the day’s cases. After 15 minutes of rounds beside the bay, the group gathers inside the on-site naval veterinary hospital to review twospecial cases.

In the last month, the veterinariansworked with two interventionalradiologists at Naval Medical Center SanDiego, the human military hospital, tocheck the lungs of two dolphins—onetreated for a bacterial abscess and theother for a fungal infection. JennyMeegan, another foundation vet, pullsthe dolphins’ CT scans up on a computermonitor and shares the human doctors’interpretation of the results: both animalsare recovering nicely.

“Having that perspective and direction oncases is so helpful,” says Eric Jensen, theNavy Marine Mammal Program’smanaging veterinarian. “Comparativeveterinary medicine is not new. But theconcept of ‘one health’—unitingveterinary medicine and work in the

human health fields—has been gaininggreat momentum over the last four or fiveyears,” he notes. “And unlike youraverage veterinary clinic, we get toregularly work with specialists in thehuman field, thanks to our access to Navyand Army physicians. Those resourcesreally elevate our wellness program.”

For the CT scans, the dolphins traveled tothe human naval hospital and underwentthe procedure just like human patientsdo. “Our animals are trained tovoluntarily beach and be transported allover the world,” says Jensen. “So theyhave no problem going into a humanhospital—though they do tend to draw abit of a crowd. We just roll them downthe hall [on a gurney], lay them down [onthe plastic-covered CT machine] andclean up thoroughly afterwards.” Toprevent overheating, the dolphins arecontinually kept wet with sponges orspray bottles.

in addition to providing direct care to theNavy marine mammals, foundationveterinarians also work on specialprojects to solve particular clinicalproblems in marine mammal medicine orto research disease pathology.

Smith and Venn-Watson, for example,have enlisted a group of experts inveterinary and human medicine from theUniversity of Texas Southwestern,University of California, San Diego,Dolphin Quest and SeaWorld San Diegoto determine why dolphins develop

Above: Dolphins undergo a physical.


Tufts University

kidney stones. The project was inspired byRake, a geriatric male dolphin that wentinto renal failure five years ago.

While creating a treatment plan for Rake,“we talked to other [marine mammal]facilities and people working with wildanimals to see if kidney stones had similarlyaffected any dolphins they’d seen,” saysSmith. “As soon as we realized it was ahealth problem impacting many dolphins,it became a collaborative effort to learnwhy, and how to prevent and treat it.”

The multidisciplinary research team sincehas discovered that low levels of citrate inthe urine may be a risk factor for kidneystones in dolphins—and that appears tobe the case in humans, too. They are nowworking on pinning down the cause of thiscondition, known as hypocitraturia, withan eye toward developing a treatment for dolphins.

The similarities between dolphins andhumans mean that many advances indolphin medicine could influence howphysicians understand and treat humandiseases. Venn-Watson, a veterinaryepidemiologist who holds an MPH fromEmory University, said the informationcontained in the Navy’s marine mammaldatabase significantly surpasses thebreadth of any population study sheencountered during her two yearstraveling the world as a project directorfor the World Health Organization’sGlobal Foodborne Infections Network.Even a seemingly routine study can yieldunimaginable results.

Consider this simple exercise: Venn-Watson compared more than 1,000fasting and post-feeding blood samplestaken from 52 dolphins over seven years.She was surprised to find that the dolphindata did not match similar studies in otheranimals. Instead, the blood changes inthese dolphins mimicked those seen inlarge-scale studies of people with type 2 diabetes.

We humans need plenty of glucose, asugar transported through the blood, tofeed our big brains. The hormone insulinhelps our bodies regulate the metabolismof carbohydrates and fats. People with type2 diabetes either do not produce enoughinsulin or are immune to insulin’s effects,promoting the buildup of too much glucosein their blood, a condition known as insulinresistance. This blood sugar overload canlead to severe health problems, includingheart disease, stroke, blindness, nervedamage and kidney failure.

Like humans, dolphins have large bloodsugar demands because of their largebrains. But unlike humans, insulinresistance in dolphins may beadvantageous.

“Amazingly, dolphins are diabeticlike whenthey need it and non-diabetic when theydon’t,” explains Venn-Watson. Dolphinsseem to activate insulin resistance,causing temporarily high blood sugarduring short overnight fasts, she says.They then revert to a non-insulin-resistantstate as soon as they eat a meal.

“If we can find and figure out how to flipthat switch off and on in humans, thatcould be a great benefit to people withdiabetes,” she says.

Venn-Watson believes that dolphins mayhave evolved this way to stretch the limitedstores of glucose found in their extremelyhigh-protein, low-sugar diet, whichconsists entirely of fish. That notion issupported by research that Ridgwayconducted in the 1970s. When dolphinswere fed sugar, they had high glucoselevels that lasted up to 10 hours. Thosestudies showed that dolphins’ diabetes-like systems do not have the ability tohandle high-sugar meals.

The foundation is now working with thenation’s leading research institutes,including the Salk Institute for BiologicalStudies in La Jolla, Calif., to identify that

genetic on/off switch in hopes ofeventually testing a diabetes cure in miceand then in humans.

This one-health strategy is catching on. Lastwinter, the National Marine MammalFoundation welcomed 40 scientists fromaround the country, more than half of themfrom human medicine, to brainstorm newresearch projects. The gathering generateda five-year strategy and a prioritized list ofstudies in geriatric health, metabolicdiseases and infectious diseases.

“My job is to take that research road mapand bring it to life,” says Venn-Watson.Her typical work week involves reachingout to potential collaborators from thehuman medicine side. The good news isthat many are eager to invest their timeand expertise in work that could advanceboth dolphin and human health.

For example, foundation veterinarians andhuman medicine researchers arecollaborating to assess whether changessuch as mild chronic inf lammation, highcholesterol and decreasing muscle massseen in aging dolphins—which mimicchanges seen in aging humans—areassociated with particular health problemsand if targeted therapeutics can improvethe quality of life in their golden years.

“That kind of breakthrough would be awin-win for animals and humans,” saysVenn-Watson. “This is one health. Bycaring for one species, we can care for many.”

Cynthia Smith, V99, examines a dolphin’s CT scan; behind her is the veterinary epidemiologist StephanieVenn-Watson, V99. Mauricio Solano, head of diagnostic imaging at the Cummings School, helped Smithdevelop CT, ultrasound and X-ray techniques for the Navy marine mammals.

IMAGINE A LANDSCAPE WITH PEAKS ANDvalleys, folds and niches, cool, dry zones andhot, wet ones. Every inch is swarming withdiverse communities, but there are no cities,no buildings, no fields and no forests.

You’ve probably thought little about theinhabitants, but you see their environmentevery day. It’s your largest organ—your skin.“The skin is like our shell. That’s what peoplesee of us first,” says Elizabeth Grice, who justfinished a postdoctoral fellowship in geneticsat the National Institutes of Health (NIH) inBethesda, Maryland. “It’s a defining feature,but it’s also an important organ for humanhealth.”

Our skin is home to about a trillionmicroscopic organisms like bacteria andfungi. Together, these creatures and theirgenetic material—their genomes—up themicrobiome of human skin.

Grice studies the skin microbiome to learnhow and why bacteria colonize particularplaces on the body. Already, she’s found thatthe bacterial communities on healthy skin aredifferent from those on diseased skin.

She hopes her work will point to ways oftreating certain skin diseases, especiallychronic wounds.

Collage photographs of body parts in fouradjacent hexagonsCollage photographs ofbody parts in four adjacent hexagonsCollagephotographs of body parts in four adjacenthexagonsCollage photographs of body partsin four adjacent hexagons

“I like to think that I am making discoveriesthat will impact the way medicine ispracticed,” she says.

Entering the FieldGrowing up in Wisconsin and Iowa, Gricewas exposed to biology at a young age—butin a field, not a laboratory.

“My first job was detasseling corn,” sheremembers. Pulling the tassel, orpollenproducing flowers, off the tops of cornplants is a way to breed highyield hybrid cornwith specific traits.

Summer days in the fields were hot andtaxing. “That was when I realized I didn’t wantto do manual labor,” Grice laughs.

When Grice was in middle school, hermother went back to college for a bachelor’sdegree in biology. Reading off flashcards tohelp her mom study sparked Grice’s owninterest in science.26 ResearchSaves.org Spring/Summer 2013

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In high school, Grice trained to become acertified nursing assistant and worked in anursing home. Then she enrolled at LutherCollege in Decorah, Iowa for a bachelor’sdegree in biology, with dreams of being adoctor.

When biology professor Marian Kaehlerannounced a summer researchopportunity for seasoned students,Grice—a freshman with no labexperience—knocked on Kaehler’s door10 minutes later and asked for the job.

“She was determined, enthusiastic andconfident, and we decided to try it,”Kaehler remembers. “It worked outextraordinarily well.”

Grice studied plant genetics in Kaehler’slab throughout college. She found theenvironment, with its experiments andchallenges, a more comfortable fit than acareer focused on seeing patients—orsummers breeding corn.

Several research internships later, Griceearned a Ph.D. in human genetics andmolecular biology from the Johns HopkinsSchool of Medicine before coming to NIHto tackle bacterial genomics.

The Good, the Bad and the AcneWhen you use antibacterial hand soap ortake antibiotics, it’s easy to think ofbacteria as bad guys. After all, Salmonellaand E. coli can give you food poisoning,and Staphylococcus aureus (S. aureus)can cause pneumonia, meningitis orserious wound infections.

But bacteria aren’t all bad. Many areharmless, and some are actually veryhelpful. On the skin, Staphylococcusepidermidis protects us by taking up space that the harmful S. aureus wouldotherwise colonize.

The common skin bacterium that causesacne works the same way. “It’s occupyinga niche so that other, more potentiallyharmful bacteria don’t invade,” Grice explains.

It might sound unhealthy or evendangerous to have skin that’s teemingwith bacterial colonies. But as Gricepoints out, it’s completely ordinary.

Your skin was sterile only once in yourlife—when you were in the womb. Minutesafter you were born, bacteria began to colonize it. Your body relies on some of these bacteria as part of its first line of defense.

Many bacteria on the skin defend themselves by secreting antimicrobial peptides, or small proteins that kill harmful invaders. In protecting themselves, they also protect us.

Diverse SettlementsLike plants, bacteria don’t all fare well in the same environment. Some are better suited to moist, humid folds like the armpit or navel. Others colonize dry expanses like the forearm or oily nooks like the side of the nostril.

Grice has surveyed the microbiallandscape of human skin like atopographer charts a territory and ananthropologist studies its populations.

From a study of 20 different skin sites on agroup of healthy people’s bodies, Griceand her colleagues identified three types ofenvironments: moist, dry and sebaceous(oily). Then they investigated which types ofbacteria colonize what sites.

Scientists have traditionally studied skinbacteria by smearing a sample of themonto a layer of nutrientrich gel in a Petri dish.

But 99 percent of the microbes won’t growon laboratory plates, because they needto interact with other members of theskin’s bacterial community to survive. It’salso tough to replicate the exact nutrientsand environment the skin provides.

Grace calls this “the great plate countanomaly”—bacteria that grow well in thelab aren’t necessarily major players on the skin.

Grice employed a newer technique thatuses a gene called 16S rRNA.

This gene provides the code for part of abacterial ribosome, the essentialmachinery needed to make proteins.

The 16S rRNA gene is present in everyknown bacterium, but in each one, it hasa slightly different DNA sequence.Scientists can use the sequence of thisgene to classify the bacteria.

The Petri dish method has uncovered 10different types of skin bacteria. Themethod Grice used revealed more than athousand. Her study was the first to usethe technique for such a large survey ofhuman skin.

She found that moist areas tend to hostsimilar bacterial communities in all of hervolunteers. The same holds for dry andsebaceous areas. Each skin environmentdetermines its bacterial inhabitants just asan outdoor environment determines itsplant life—rainforests support leafy trees,while deserts have cacti.

Even with these patterns, the skin still hasa surprising amount of variation fromperson to person. ➤

Body BacteriaExploring the Skin’s Microbial Metropolis BY ALLISON MACLACHLAN


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Skin microbiomes are like snowflakes: Notwo are exactly alike. Your unique patterndepends on things like your age, sex, sunexposure, diet, hygiene and even whereyou live and work.

Microbes in MedicineBy getting a sense of bacteria on healthyskin, Grice hopes to figure out what’sdifferent about the microbes on diseasedskin—and maybe even find a way to fixthe problem.

She’s most excited about applying herwork to the chronic wounds that arecommon in people who have diabetes orspend most of their time in beds orwheelchairs.

People with diabetes can lose some ofthe sensation in their limbs, making itharder for them to feel pain and easierfor any of their injuries to fester.

On top of that, they may have poorblood flow, which makes healing tough.

As Grice explains, your body needs bloodto deliver oxygen, immune cells andimportant proteins to the site of an injuryto help cells regenerate.

A Problem AfootAlmost 10 percent of the United Statespopulation has diabetes, and up to aquarter of these 24 million people willget a painful wound known as a diabeticfoot ulcer.

These ulcers are very difficult andexpensive to treat. And the problem isincreasing: As obesity rates rise,diabetes—and diabetic foot ulcers— arebecoming more common.

“It’s such a farreaching problem that it’sclearly an area of need,” says Grice.“That’s what really drives me the most.”

Grice suspects that bacteria make chronicwounds worse because they spur thehuman immune system to triggerinflammation. Although designed to killinfected cells, inflammation also preventsskin cells from regenerating after an injury.

The immune system acts slightlydifferently in each of us, thanks to ourgenetics. Grice’s work takes a microlevellook at interactions among human genes,the immune system and the skin’sbacterial communities.

Defense MechanismsTo investigate what role bacteria play indiabetic wounds, Grice used a group oflaboratory mice bred to display commonfeatures of diabetes—like wounds thatdon’t heal well.

Grice and her colleagues took skin swabsfrom both diabetic and healthy mice, andthen compared the two. Using the 16SrRNA technique, they found that diabeticmice had about 40 times more bacteriaon their skin, but it was concentrated intofew species. A more diverse arrabacteriacolonized the skin of healthy mice.

“People with diabetes have high bloodsugar, which is known to change theskin’s structure,” says Grice. “Thesechanges likely encourage a specificsubset of bacteria to grow.”

The researchers then gave each mouse asmall wound and spent 28 daysswabbing the sites to collect bacteria andobserving how the skin healed.

They found that wounds on diabetic micestarted to increase in size at the same timeas wounds on healthy mice began to heal.

In about 2 weeks, most healthy micelooked as good as new. But most diabeticmouse wounds had barely healed evenafter a month.

Interestingly, bacterial communities in thewounds became more diverse in bothgroups of mice as they healed—althoughthe wounds on diabetic mice still had lessdiversity than the ones on healthy mice.

“Bacterial diversity is probably a goodthing, especially in wounds,” says Grice.“Often, potentially infectious bacteria arefound on normal skin and are kept incheck by the diversity of bacteriasurrounding them.”

Then Grice and her colleagues examineddifferences between healthy and diabeticmice at the genetic level. They focusedon the genes that control aspects of theimmune system in the skin.

They found distinctly different patterns ofgene activity between the two groups ofmice. As a result, the diabetic mice putout a longerlasting immune response,including inflamed skin. Scientists believeprolonged inflammation might slow thehealing process.

Grice’s team suspects that one of the maintypes of bacteria found on diabeticwounds, Staphylococcus, makes one of theinflammationcausing genes more active.

Now that they know more about thebacteria that thrive on diabetic wounds,Grice and her colleagues are a stepcloser to looking at whether they couldreorganize these colonies to help thewounds heal.

Bacterial diversity is probably a good thing, especially in wounds…

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More Than Skin DeepSkin isn’t the only place in the body that’scrawling with bacteria.

Grice also spends time studying bacteriathat live in the intestines. There too,microbes can be helpful.

Certain strains of E. coli in our digestivetracts help keep dangerous bacteria atbay and produce Kand Bcomplexvitamins, which our bodies can’t makeenough of on their own.

Grice is involved with a study ofHirschsprung disease, a genetic disorderthat leaves parts of the digestive tractwithout enough nerve endings to pushwastes out.

Some children born with the disease getenterocolitis, a painful inflammation in thegut, and others don’t. Together withgeneticist Bill Pavan, who also works atNIH, Grice is looking at gut bacteria tosee if their distribution differs between thetwo groups. If the researchers find apattern, it might help predict whichpatients will need surgery to reduceinflammation. Grice and Pavan also thinkthat redistributing some of the bacteria ininflamed intestines might help. Pavanadmires Grice’s confidence anddedication to her science, and he alsosays that working with her is a lot of fun.“She is driven to get highquality researchdone, but she’s still extremely friendly andinteractive on a personal level,” he says.“She has an infectious laugh.”

Pavan said Grice is well known forwhipping up impressive treats likeminiature chocolate mice, which are verypopular in the lab. And whenever alabmate has a birthday, Grice brings in acustombaked cake with whatever flavorand frosting the person wants.

“Most people wouldn’t suspect that I’mvery domestic,” says Grice, who listscooking as one of her hobbies. “You getto a point where you’re comfortableexperimenting with recipes and seeingwhat works.”

Grice likes getting creative with herexperiments in the kitchen as well as in thelab. “My husband doesn’t really eatvegetables, so it’s always a challenge towork around that,” she laughs.

Taking Exploration GlobalFor Grice, exploring diverse landscapesand populations goes far beyond skinsamples. Outside of her work, she enjoystraveling to exotic locations to soak up the culture.

She and her husband were married inBelize, a country they chose for its naturalbeauty and its preserved culture. “It’s oneof those places that you feel isn’t overrunby civilization,” she says.

Highlights included exploring Mayanruins, relaxing on beaches and snorkelingin the striking coral reefs off the coast.

Grice also counts Greece among herfavorite destinations because of itsarchitecture and the laidbackMediterranean attitude. “I love Athens andall the old ruins that are just integratedinto the city,” she says.

When she’s home, Grice likes to exploreother cultures and civilizations by reading.A selfprofessed bookworm, her favoritegenre is historical fiction, including novelsabout the Tudor period in Britain.

Tying her hobbies to her career choice iseasy for Grice. “I really like experiencingdifferent cultures, and science is somulticultural—you get to interact with adiverse group of people,” she says.

Charting New GroundDuring the preparation of this article,Grice was considering job offers for afaculty position. She decided to join theUniversity of Pennsylvania’s dermatologydepartment and will start working there inJanuary 2012.

In her new job, she will continue herresearch on the wound microbiome andteach graduate and medical students.

She hopes that she, like her longtimementor Marian Kaehler, will inspire andchallenge her students.

“She was just so tough, and I reallyrespected that,” Grice says of Kaehler.

“Having a female mentor was also reallyimportant to me, because otherwise, howdo you picture yourself in that role?”

Even now that she’s landed that role,Grice’s ambition isn’t flagging. She aimsto sustain a successful research program,improve the way chronic wounds aremanaged and keep time for personalgoals like traveling to new continents.

Kaehler, for one, is confident that Gricewill succeed. “She has a very strong senseof self, and there’s nothing moreimportant for people making careerdecisions than knowing where you’regoing to find a niche that makes yousatisfied and challenged,” she says.

Like the bacteria she studies, Grice knowswhere she thrives.

Skin isn’t the only place in the body that’s crawling with bacteria.

Grice loves to experience the natural beauty and local culture in countries like Belize, Greece and Costa Rica.

– Photo by ELIZABETH GRICE

We are dedicated to the advancement of COMPASSIONATE CARE

for laboratory animals through EDUCATION, TRAINING and consistent INFORMATION EXCHANGE

for the benefit of human and animal health.

TEACHING THE HIGHEST STANDARDS OF ANIMAL CARE AND USE

www.lawte.org

TheCareer Day Kit is a coord inated, nation-wide endeavor designed to educateelementary school students about humane

and responsible animal research. According to the U.S.Department of Education, there are more than 130,000elementary and secondary schools in the U.S. If scientistsacross the country can make one 20-minute presentation inone classroom in every one of these 130,000 schools, wehave the potential to reach 2.6 million students in just oneyear! It is possible to have a HUGE impact by simply takingtime out of the day to teach children about animal research.

The Career Day Kit empowers you to make effective presentations in elementary school classroomsthat will educate children about science careers and animal research. You will help illuminate theimportance of animal research and have an impact on their thinking for their entire lives. It isimportant to reach children early and often with the truth about biomedical research. The future ofscience and human health is at stake and you can help!

The Foundation for Biomedical Research (FBR) Career Day Kit includes a DVD with lab footage,stickers for the children, a presenter’s “how to” guide, and several useful props.

ORDER YOUR CAREER DAY KIT TODAY…ONE FREE KIT PER INSTITUTION. To order, please contact Nahla at 202-457-0654 or send her an email at [email protected]


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Mice without the gene for myostatin (right)have nearly twice as much muscle mass asnormal mice (left). Working with mice, Johns Hopkins researchers have solved a keypart of a muscle regeneration mystery plaguing scientists for years,adding strong support to the theory that muscle mass can be builtwithout a complete, fully functional supply of muscle stem cells.

“This is good news for those with muscular dystrophy and othermuscle wasting disorders that involve diminished stem cellfunction,” says Se-Jin Lee, M.D., Ph.D., lead author of a report onthe research in the August issue of the Proceedings of the NationalAcademy of Sciences and professor of molecular biology andgenetics at the Johns Hopkins University School of Medicine.

Muscle stem cells, known as satellite cells, reside next to musclefibers and are usually dormant in adult mammals, includinghumans.

After exercise or injury, they are stimulated to divide and fuse,either with themselves or with nearby muscle fibers, to increase orreplace muscle mass. In muscle wasting disorders, like musculardystrophy, muscle degeneration initially activates satellite cells toregenerate lost tissue, but eventually the renewal cycle isexhausted and the balance tips in favor of degeneration, theresearchers explain.

Muscle maintenance and growth under healthy, non-injuryconditions have been more of a mystery, including the role ofmyostatin, a protein secreted from muscle cells to stop musclegrowth. Blocking myostatin function in normal mice causes themto bulk up by 25 to 50 percent. What is not known is which cellsreceive and react to the myostatin signal. Current suspects includesatellite cells and muscle cells themselves.

In this latest study, researchers used three approaches to figure outwhether satellite cells are required for myostatin activity. They firstlooked at specially bred mice with severe defects in either satellitecell function or number. When they used drugs or geneticengineering to block myostatin function in both types of mice,muscle mass still increased significantly compared to that seen inmice with normal satellite cell function, suggesting that myostatinis able to act, at least partially, without full satellite cell function.

Second, the researchers guessed that if myostatin directly inhibitsthe growth of satellite cells, their numbers should increase in the

absence of myostatin. The researchers marked the satellite cellswith a permanent dye and then blocked myostatin activity with a drug.

Mouse muscle mass increased significantly as expected, but thesatellite cells did not increase in number, nor were they foundfusing with muscle fibers at a higher rate. According to Lee, theseresults strongly suggest that myostatin does not suppress satellitecell proliferation.

Third, to furtherconfirm their theorythat myostatin actsprimarily throughmuscle cells and notsatellite cells, the teamengineered mice withmuscle cells lacking aprotein receptor thatbinds to myostatin. Ifsatellite cells harbormost of the myostatinreceptors, removal ofreceptors in musclecells should not alter myostatin activity and should result inmuscles of normal girth. Instead, what the researchers saw was amoderate, but statistically significant, increase in muscle mass. The evidence once again, they said, suggested that muscle cellsare themselves important receivers of myostatin signals.

Lee notes that, since the results give no evidence that satellite cellsare of primary importance to the myostatin pathway, even patientswith low muscle mass due to compromised satellite cell functionmay be able to rebuild some of their muscle tone through drugtherapy that blocks myostatin activity.

“Everybody loses muscle mass as they age, and the most popularexplanation is that this occurs as a result of satellite cell loss. If youblock the myostatin pathway, can you increase muscle mass,mobility and independence for our aging population?” asks Lee.“Our results in mice suggest that, indeed, this strategy may be away to get around the satellite cell problem.”

Authors on the paper include Se-Jin Lee, Thanh Huynh, Yun-Sil Lee andSuzanne Sebald from The Johns Hopkins University, Sarah Wilcox-Adelman of Boston Biomedical Research Institute, Naoki Iwamori andMartin Matzuk of Baylor College of Medicine, and Christoph Lepperand Chen-Ming Fan from the Carnegie Institution for Science.

Hopkins ResearchersSolve Key Part of

Old Mystery in Generating

Muscle MassImplications for treating muscular dystrophy and other muscle wasting diseases

BY VANESSA MCMAINS, PH.D.

–Photos by S

E-JIN

LEE LAB

Everybody losesmuscle mass as theyage, and the mostpopular explanationis that this occurs asa result of satellitecell loss....


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collaborationcould someday

help dogs and

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WhenOlga Troyanskaya’s dog Jessy fell ill in early2006, the vet had painful news. “The doctor

told me she had less than a week to live,” Troyanskaya said.

Troyanskaya sought a second opinion with a dog cancerspecialist, with an unexpected result: The appointmentlaunched an innovative research collaboration to learn moreabout cancer, possibly leading to new treatments for dogs likeJessy and humans as well.

At the appointment, Troyanskaya, a computational biologist atPrinceton University, got to talking with Jessy’s canineoncologist, Karin Sorenmo, head of the small animal oncologyservice and an associate professor of oncology at the RyanVeterinary Hospital of the University of Pennsylvania.

The two researchers discovered that they share a goal to learnabout how to understand and treat cancer. Troyanskaya wasinterested in the genes involved in cancer; Sorenmo wanted toexplore the mechanisms of cancer to find new ways to treat herpatients, most of them well-loved pets.

“A collaboration seemed like a unique way to look at thequestion of cancer progression,” said Troyanskaya, anassociate professor of computer science with a jointappointment in the Lewis-Sigler Institute for IntegrativeGenomics at Princeton.

It is also a way to help dogs like Jessy, she said. For years,Jessy was Troyanskaya’s canine collaborator, accompanyingthe scientist to the lab where she’d station herself next to herowner’s desk.

Relying on a comparative approachAfter months of conversations while Jessy was treated withchemotherapy, Troyanskaya and Sorenmo started acollaboration between their two laboratories, which arelocated about 50 miles apart.

Sorenmo provides samples of tumors that she has extractedfrom her canine patients during surgery and Troyanskayaanalyzes the genes in the tumors.

All of Sorenmo’s patients developed cancer through naturalprocesses, just as humans do. She treats them with the sametypes of therapies humans receive, including radiation,chemotherapy and surgery. She and her colleagues examineeach extracted tumor, noting its size and other attributes, butnow with the Princeton collaboration, Sorenmo can learnmuch more about the cancers she treats.

The study of naturally occurring cancer in animals, and itsapplication to human cancer, is called “comparativeoncology.” Sorenmo is a leader in this emerging field.Comparative oncologists recognize that much can be learnedabout cancer by comparing animals and humans.

“Dogs get all the same cancers that humans get,” said ChandKhanna, director of the Comparative Oncology Program at theCenter for Cancer Research, which is part of the U.S. NationalCancer Institute. “With dogs, we can ask many questions thatone cannot ask in mouse preclinical models of cancer andcannot answer in human clinical trials.”

Troyanskaya and Sorenmo are looking for answers by studyinga type of cancer that poses a high risk for both dogs andhumans: mammary cancer.

Mammary tumors are the most common kind of tumor infemale dogs that have not been spayed. Breast cancer is themost commonly diagnosed cancer in women, striking one ineight women during their lifetime.

Yet not all dogs or women succumb to the disease: In someindividuals, the tumors are benign and grow slowly over years,while in others the cancerous cells grow rampantly andeventually kill. Troyanskaya and Sorenmo hope to learn moreabout why this is so.

Computational geneticist Olga Troyanskaya of Princeton University (left) and KarinSorenmo, a veterinary oncologist at the University of Pennsylvania Ryan VeterinaryHospital, pose with Sorenmo’s dog Ruby. The two scientists are collaborating on astudy to explore genetic factors contributing to mammary cancer growth in dogsbeing treated at the hospital, with the goal of enhancing the understanding ofcancer in both dogs and humans. – Photo courtesy of OLGA TROYANSKAYA


Princeton University

BY CATHERINE ZANDONELLA

Canines are helpful in the study of mammary cancer for anotherreason: Each female has eight to 10 mammary glands, making itpossible to study several tumors — each arising separately fromthe other and therefore genetically unique — in one individual.Studying separate breast tumors in humans is usually not possiblebecause it is rare for a woman to develop more than onespontaneous tumor in the breast.

By studying different tumors isolated from a single dog, theinvestigators can hone in on gene signatures related to tumordevelopment without having to subtract out gene signatures fromthe rest of the body. “In humans, a lot of times the variability is solarge across individuals that it completely masks any signal ofprogression-related variability between the tumors,” saidTroyanskaya. “With dogs, you can look at tumors at differentstages of cancer progression in the same genetic background,within each individual.”

Evaluating genetic signaturesAt Princeton, Troyanskaya’s team has begun evaluating the tumorsamples to look for genetic signatures that may be correlated tothe progression of mammary cancer in dogs. This type of genome-wide search became possible with the sequencing of the caninegenome in 2005.

To evaluate the genes in the tumors, Troyanskaya’s group, whichincludes molecular biology graduate student Dmitriy Gorenshteyn,isolates the genetic material in the tumor samples. Theinvestigators look at which genes are turned on, or “expressed,”and which are turned “off.” They try to tie the resulting geneexpression pattern to tumor attributes such as rapid growth.

Several studies of gene expression patterns have been conductedfor human breast cancer, but this is possibly the first such study ofcanine mammary tumors. Nor have comparisons between thegene expression patterns of human and dog tumors been made.

The dog genome provides special challenges because dog breedshave different genetic variations. Using statistical methods, theresearchers have already started identifying the canine equivalentsof human genes based not only on their sequence, but also ontheir functions in the body. “The vast majority of dog genes have acorresponding human gene,” Troyanskaya said.

An expert in combining computer science with biology, Troyanskayahas developed computer programs that can sort through the datagenerated by the genome-scale gene expression studies to detectpatterns of gene expression that correlate with increased tumorsize. The team, which includes quantitative and computationalbiology graduate student Jonathan Goya, plans to follow up thegene expression studies with a search for other abnormalities suchas extra copies of genes, known as copy number aberrations, andchanges in protein levels. Eventually the group hopes to linkgenetic pathways to the progression of a tumor from a harmlessovergrowth of cells to a deadly spreading malignancy.

Providing care to shelter dogsAt the beginning of the collaboration, Sorenmo collected samplesof tumors that she surgically removed from Penn vet clinic patients,typically pet dogs from comfortable homes. But it nagged at herthat homeless dogs were not able to get similar care.

Stray dogs are also the least likely to have been spayed, and theresulting hormone levels put them at much greater risk thanspayed animals of developing mammary tumors.

To help homeless dogs, Sorenmo started the Penn Vet ShelterCanine Mammary Tumor Program in 2009 to provide cancertreatment to dogs living in shelters. “The shelter program is a wayto provide high-quality care for some of the neediest dogs, whilehelping to further our knowledge of both canine and humanbreast cancer,” Sorenmo said.

Under the program, which is funded by family foundations, shelterdogs are treated for cancer free of charge. Dogs that are lateradopted continue to obtain follow-up treatments at no cost to theowner. Lisa Hertzog, a resident of Reading, Pa., has adopted twodogs participating in the program. “It is really a rewardingexperience,” she said. “The dogs are really loving, and they giveback so much more than what you give them.”

Since beginning the collaboration with Troyanskaya, Sorenmo andher colleagues at Penn have operated on more than 60 shelterdogs. Studies of these samples are already starting to reveal factorsthat may help clinicians predict the progression of the tumors.

Gaining support from dog loversDespite its advantages, comparative oncology — which involvespets rather than laboratory animals — is not yet a mainstreamapproach to studying human cancer. To fund her initial studies,Troyanskaya received funding from Princeton’s Project X, a fund setup by businesswoman and philanthropist Lynn Shostack to supportpioneering and speculative projects.

Further help came this January from 2 Million Dogs, a newfoundation dedicated to finding cures for canine cancer. Itsfounder, Luke Robinson, walked 2,000 miles from Austin, Texas, toBoston in 2008 to raise awareness of canine cancer and build agrassroots network of dog lovers and cancer researchers. He metSorenmo on his walk through Philadelphia, and the foundationawarded its first grant to the collaboration between Sorenmo andTroyanskaya. In January, 2 Million Dogs presented a $50,000check to the researchers to help them purchase the laboratorysupplies for the study of genes involved in canine cancer.

“This work is incredibly important to anyone who has had a lovedone, whether dog or human, who has had cancer,” said Robinson,whose dog Malcolm died of cancer.

Through the work funded by 2 Million Dogs, Troyanskaya and herteam hope to find gene expression patterns that govern thetransformation of a tumor from a benign to malignant state,contribute to tumor growth and govern metastasis. Theinvestigators anticipate that their studies will be a starting point fordeveloping diagnostic methods that veterinarians and doctors canuse to predict whether a newly discovered tumor will grow slowlyor rapidly. They also hope to identify novel pathways that couldserve as targets of new drugs to treat cancer.

Troyanskaya’s faithful laboratory assistant and canine companionJessy succumbed to cancer six months after beginning hertreatment. But Troyanskaya is optimistic that the collaboration thatemerged from Jessy’s illness will provide a lasting legacy. “Wehave the potential to contribute new findings that could lead tobetter cancer diagnosis and treatments for humans and for ourdogs,” she said.


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OSU RESEARCH COULD HELP ‘MANY DOGS AND CATS LIVE A LONG, HEALTHY LIFE’

An OSU educator could have a profound impact on diagnosing and treating a fungal disease, which has become pervasive in Oklahoma.

Andrew Hanzlicek, DVM, M.S., diplomate of the American College of Veterinary Internal Medicine in small-animal internal medicine, is an assistant professor of small animal

internal medicine at OSU’s Veterinary Hospital. His research on histoplasmosis — a fungal disease endemic in Oklahoma — may change the way the disease is diagnosed and treated.

— by DERINDA BLAKENEY

TACKLING A FUNGAL DISEASE


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Histoplasma capsulatum, a soil-borne fungus, is found in temperate and subtropical regions throughout the world. In the U.S., histoplasmosis appears in the Ohio, Missouri and Mississippi river valleys as well as in Oklahoma. The disease is a common systemic fungal infection of many dogs and cats inOklahoma and occurs when microconidia — the mycelial form of Histoplasma sp. found most abundantly in nitrogen-rich soil — is inhaled or sometimes ingested.

“I saw this disease when I worked at Texas A&M University and at Kansas State University, but Iwas surprised at how frequently we see it here at OSU’s Veterinary Hospital,” says Hanzlicek.“Because of its small size, the microconidia can penetrate deeply into the lungs when inhaled.Histoplasmosis is a disease that can affect the respiratory system, the gastrointestinal tract or theskin in cats and dogs. Sometimes it affects the bone marrow, eyes, the brain — it can go virtuallyanywhere in the body.”

Common clinical signs include lethargy, weight loss, anorexia and fever unresponsive toantibiotics. An infected animal — especially a dog — may have diarrhea.

Traditionally, the diagnosis of histoplasmosis is made from clinical signs and finding fungalorganisms from affected tissue or fluid samples and, in some cases, a fungal culture.

“Fungal culture has the disadvantages of lacking sensitivity, requiring specialized laboratoriesand having a slow turnaround time (weeks).”

Currently, Hanzlicek uses a test from MiraVista Diagnostics in Indianapolis. The test can beperformed on body fluids of the potentially infected animal.

“We can use urine samples, blood samples or fluid from a lung wash,” explains Hanzlicek. “If a protein from the cell wall is in the sample, the dog or cat will test positive for histoplasmosis.”

Hanzlicek started a clinical trial in June 2012 to determine how treatment with antifungaltherapy affects the antigen test.

“From previous information in over 70 dogs and 30 cats, we are convinced this test is accurate,and in some cases, this test has changed the way we diagnose the disease,” he says. “Beforethis test, we diagnosed histoplasmosis based on clinical signs and finding the fungus throughinvasive tissue biopsy or needle aspirate procedures. Now it may be as easy as submitting aurine or blood sample. Next, we need to find out if it can also be used to monitor or guideantifungal therapy.

“Right now, the test is used to monitor treatment in humans. We don’t have data on animals,and that’s why we are doing the study,” saysHanzlicek. “When you test positive forhistoplasmosis, the test readout gives youa number. For example, it could be 8.2.You treat the person and then retest. Theresult has to be below a certain numberbefore you stop treatment. If you stopantifungal therapy too soon, the infectioncould return. If the test result decreaseswith treatment like we expect, it shouldwork the same for animals as it does forhumans.”

He has applied for grant money to coverthe research and hopes to have 20 dogsand 20 cats known to be infected withhistoplasmosis in the program.

“We will monitor how the antigen testchanges during treatment, which will helpus decide if we can use the test tomonitor treatment. I anticipate it will take12 to 18 months to gather the data,” he says.

Dr. Andrew Hanzlicek examines Midget,owned by Lois Crain of Ringling, Okla., withDr. Jennifer Chang, a resident in small-animalinternal medicine.

TACKLING A FUNGAL DISEASE

“If we can diagnose histoplasmosisearly on and if we treat for theappropriate amount of time, theanimals have a pretty good chanceof making it. This isn’t an end-of-lifedisease. It affects young, otherwisehealthy dogs and cats. If we candiagnose it with a simple fluid testand monitor treatment withoutinvasive procedures, we have abetter chance of helping many dogsand cats live a long, healthy life.”


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In 2008,on the GrandCanyon’s

southern rim, a biologist named Eric York found a dead mountainlion with a bloody nose but no othersigns of trauma. He took it back tohis garage to perform an autopsy,which revealed nothing unusual.

Two days later, York developed abad cough. He felt weak, achy,tired. His doctor told him he had aflu-like illness and sent him home.Two days after that, York was dead.

This time, the autopsy did revealsomething. York was stricken withthe plague, also known as the BlackDeath, the same disease that wipedout half of Europe during thefourteenth century. Public-healthofficials gave antibiotics to everyonewho had come in contact with York.

No one else died. Disaster averted.But how did York’s doctor misssomething as uniquely horrifying as the plague?

Turns out just about every doctor would’vemissed it, according to UNC’s BillGoldman. “The first symptoms of theplague really are indistinguishable fromthe flu,” he says. But unlike the flu, theplague is already well on its way toshutting down the lungs by the time apatient begins to feel sick. It’s a sneaky,extremely contagious, and fatal disease,three reasons why governments andresearchers think the plague is abioterrorism threat—a twenty-first-centuryweapon of mass destruction.

In medieval times of war, combatantswould catapult infected bodies over citywalls. Today, a bioterrorist attack wouldbe stealthier and a lot more dangerous.

After the anthrax scare of 2001, the U.S.government pushed for scientists toresearch various biological warfare threats,such as Yersinia pestis, the bacterium thatcauses the plague. “I hate to put it this way,but terrorists aren’t going to unload abunch of rats or fleas into town,” Goldmansays. They’ll culture the bacteria in massiveamounts. “They’ll try to spread the diseaseby an aerosol,” he says.

Victims wouldn’t smell it or see it. Theywouldn’t even feel a thing at first, but thedisease would be on a rampage.Thousands of people would get sick buthave no idea they had the plague until itwas too late to save them.

The plague is such a silent killer becauseYersinia pestis doesn’t trigger the samesort of quick immune response that mostbacterial infections do. When a personcontracts the plague, the bacteria multiplyfrom a few microbes to a billion within 48hours. But for some reason the lungs—typically very good at getting rid ofundesirables—don’t respond.

In the case of Eric York, doctors had noway of distinguishing his illness from theflu. Only when symptoms worsen—vomiting, difficulty breathing, coughing upblood—does the plague give itself away.“By then, when it’s recognizable aspneumonic plague, it’s too late to treatit,” Goldman says. The lungs are overrunwith bacteria. The pulmonary system is allbut shut down. The circulatory systemcan’t deliver antibiotics into the lungs.Patients suffocate to death. They just can’tbreathe anymore.

“Here’s the question we wanted toanswer,” Goldman says. “Is Y. pestisavoiding detection, or is it actuallysuppressing the immune responses of thelung?” The answer would give his teamclues about how to make the plague lesslike the Black Death and more like the flu,at least in terms of patient prognosis.

Goldman’s samples of Yersinia pestiscame from a repository that got itsspecimens when a Colorado woman diedof the plague in 2000. She had been

BlackDeathThreatUNC’s Bill Goldman

battles the next

outbreak of the

plague before

it happens.BY MARK DEREWICZ

– photo by Mark Derewicz and Bill Goldman


University of N

orth Carolina

infected by her cat, which had probably gotten hold of aninfected rodent. These specimens are just as deadly now, which iswhy Goldman’s team was put through stringent security checksbefore being allowed to work with the organisms. The FBI hasactive files on each lab member, including Goldman.

When no one is working in the Goldman lab, sealed and lockeddoors separate humans from the containers that hold thebacteria. Lab technicians change into protective clothing in adesignated chamber between the outer lab and the inner labwhere they handle the samples. They attach to their heads adevice that continuously pushes air downward to lessen thechance that they’ll breathe in a pathogen. They open specimencontainers only under a special hood, into which they reach withgloved hands to conduct experiments.

One of the reasons Yersinia pestis is such an aggressive killer isbecause it contains a particularly nasty plasmid—a segment ofDNA that is not part of a bacterium’s chromosomes but canreplicate and transfer into other living things. Yersinia pestispicked up its deadly plasmid from some other organismthousands of years ago, Goldman says. He wondered howvirulent the bacterium would be without that plasmid, so his teamtook it out and placed a droplet of the specimen on the nose of asingle mouse. When the mouse breathed it in, the bacteria didn’tmultiply. In fact, they declined in numbers over four days.

The mouse never got sick. This proved that the plasmid isabsolutely critical for lung infection to spiral out of control.

Then Goldman’s team mixed the nonlethal strain of Yersinia withthe deadly strain and documented how they behaved in mouselungs. The deadly strain multiplied like mad, as Goldmanexpected, but so did the nonlethal strain.

In another experiment, his team documented how other, relativelyharmless bacteria responded when the deadly Yersinia strain waspresent in the lungs. “Even the harmless bacteria are able togrow really well when Y. pestis is present,” Goldman says. “Theyincrease from a thousand to between one million and ten millionorganisms in the lung.” Those once-harmless bacteria wind upaiding Yersinia in blocking the lung’s air passages.

Although Goldman and his team have indicted that loneplasmid, they’re still trying to pin down the mechanism thatallows Yersinia to change the lung into such a permissiveplayground for pathogens. And if they find that mechanism?“What I’d like to say is, ‘Oh, that will lead us to a drug,’”Goldman says. “But it depends on what the mechanism is.”

His team has already identified a Yersinia protein that helps thebacterium multiply inside the lung. “We have a patent on theidea of creating an inhibitor of that protein,” Goldman says, “but we haven’t found an inhibitor yet.”

Disabling that lone gene might be less a cure than a shield tokeep the disease from progressing so fast, which might givedoctors more time to treat patients.

“You have to figure out how to defeat the main barriers totreatment,” Goldman says. And in the case of the plague, themain barrier is the speed at which the disease takes hold. Aperson usually dies within three and a half or four days ofcontracting pneumonic plague. Goldman says that inactivatingthe protein his team has identified could keep patients alivelonger than usual, and that would give antibiotics more time towork. “If you can change the speed of the infection,” he says,“you’ve solved a major problem.

This approach wouldn’t help everyone infected with the plague. Itlikely wouldn’t have helped Eric York. But lengthening the timebetween initial infection and death could be enough to savethousands of lives after a bioterrorism attack.

“Imagine the worst-case scenarios,” Goldman says. “An aerosolreleased that exposes a lot of people at once, and no one wouldhave any idea they’ve been exposed. All of a sudden, everyone issick. Early symptoms are indistinguishable from the flu.”

In such cases, a cure would be best. A vaccine would be a closesecond. The next best thing would be to slow down the diseaseso treatment has a chance to work. “The plague is susceptible toantibiotics,” Goldman says. “Just not in that last 24 hours.”

Bill Goldman is chair of the department of microbiology andimmunology in the School of Medicine. He received funding from aNational Institutes of Health grant to the Southeast Regional Center ofExcellence for Emerging Infections and Biodefense, which isheadquartered at UNC-Chapel Hill.

Plague at a glance

✗ The plague was never eradicated; it thrives in the wild.

✗ Few humans are infected anymore.

✗ The organism that causes the plague is now abioterrorism threat.

✗ UNC’s Bill Goldman has found a new clue about whythe plague is so deadly and how to make it less so.

Bubonic or Pneumonic“The classic bubonic plague is a disease that’s in the wild all the time,” says UNC microbiologist Bill Goldman. “It’s constantly circulating between rodents and the fleas that infect them.”

When an infected rodent or flea bites a person, thebacterium Yersinia pestis spreads through the humanlymphatic system. Lymph nodes get inflamed and swell into hard nodules, causing incredible pain and eventualdeath. There is no cure. The inflamed lymph nodes arecalled buboes—thus the name bubonic plague. But whenYersinia pestis enters the bloodstream, it travels to lungsand patients develop secondary pneumonic plague, whichis very contagious. A cough or sneeze can easily transmitthe disease, and if someone catches the plague that way,it’s called primary pneumonic plague. “This is the worstform of the disease,” Goldman says. “It’s one reason whythe plague is a bioterrorism concern.” Infected patientsmight not know they have the plague, but they can stillspread it.


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rch Scholarships are awarded to three pre-veterinary students who wrote

outstanding essays on the need for lab animals in medical and scientific research

Foundation for Biomedical Research (FBR)announced it has awarded three collegescholarships to the winners of its 2012Animal Research Essay Contest, apartnership with Cal Poly Pomona. Firstplace is awarded Catherine Runion(graduating 2013), second place to MeganDucey-Hardos (2013), and third place toFiona Lair (2014). The winning essays will be published in the spring 2013 issue ofFBR’s semi-annual magazine,ResearchSaves.

“It is critical the next generation understandthe vital role humane and responsibleanimal research plays in both human andanimal medicine,” said FBR executive vicepresident Paul McKellips. “Catherine, Meganand Fiona have each demonstrated anexcellent grasp of how lab animal researchleads to new treatments, therapies and curesfor people and animals, and an exceptionalability to communicate that fact.”

Catherine Runion is a senior at Cal PolyPomona and will graduate in December with a degree in animal and veterinaryscience. Megan Ducey-Hardos is a seniormajoring in animal health science student;she hopes to become a registered veterinarytechnologist upon graduation. Fiona Lair ispursuing her bachelor’s degree in animalscience and plans to specialize in veterinaryradiology.

Students submitted essays on thefundamental role humane and responsibleanimal research plays in the advancement of human and animal health. Essays werejudged by representatives from FBR and Cal Poly Pomona. In addition to collegescholarships, FBR has also donated aCurVet™ Rat Training Simulator to Cal PolyPomona, in order for its pre-veterinarystudents to learn humane and ethicalhandling practices of rodents without the useof live animals. For more information aboutthe 2012 Animal Research Essay Contest orthe winners, please visit www.fbresearch.organd http://www.csupomona.edu/~agri/our-college/news.shtml.

Foundation for Biomedical Research Awards Three Scholarships to Outstanding Cal Poly Pomona Pre-Veterinary Students

Catherine Pabst

Dear Ingrid Newkirk,

California State Polytechnic University Pomona is ranked among the top public universities in

the western

United States and is known for its learn-by-doing philosophy. The

college of agriculture is the second

largest college on campus with about four thousand students, Being a student of the anim

al and

veterinary program myself 1 know just how important hands on experience with animals are, The goal for

most of us is to one day find a career in the animal field with the greater goal of im

proving their health,

As you are a member of PETA, I know you live your life with the goal of protecting a

nimals and giving

them a voice.

As a pre-vet student 1know how important animals are and I have always had a

passion for helping

them. However, the question then comes, how am I supposed to learn how to help

them if do

not have them to practice on or learn from? When we are first learning a new technique we have a few

life-like dolls that are equipped with a full set of veins, lungs and a

n esophagus. This is a perfect way to

learn what we are supposed to do but does not give a good repre

sentation of real life situation. A real

dog is not likely to just sit there, hand you its paw and be ok with l

etting you insert a catheter into its vein,

If we do not have live animals to practice our technique, more animals will have to go through more

pain in the long run.

Another huge department in the agriculture program at Cal Poly is its research department. With such a

huge campus we are lucky enough to have an equine research center, a she

ep and swine unit, a beef

unit and a mouse research facility. Each one of these facilities is run by highly e

ducated professors all

with a passion in their specific field, All the research done here is

performed under strict regulations that

put the care and welfare of the animal first, with most of the research working to he

lping the animal’s

themselves. It is very hard to create, improve and perfect any kind of tre

atment without a test subject to

see how it works. Not only are there professionals working on the

se experiments but there are students

as well. This provides wonderful hands on experience that looks g

reat on applications and resumes for

future careers or higher education opportunities.

In my own personal experience I have found that I learn so much more when 1 can not only see wha

t

I am trying to learn, but if 1 could touch and feel it as well. One of my favorite and most memorable

experiences that I have had in my five years as a pre-vet student w

as when I was able to go down to

the beef unit and meet a beautiful cow named plug. Plug, I’m sure was one of the hot topics with the

university and PETA due to the fact that he had a plug in his side t

hat enabled research to be done in

his rumen. For the brief hour that I spent with him I saw nothing except a happy an

d loved cow. I not

only got to visit with him but I got to experience the inside of a cow’s rumen, I was able to see, touch

and feel the movement and the bacteria at work processing the food that was passing

through. With

that one experience I was able to get a better understanding of the

complex process of the rumen. In

no way shape or form did I ever see Plug in any kind of stress; dogs show more fear getting a simple

vaccine injection than Plug showed surrounded by a class of stud

ents. That cow was loved by all and

received so much loving attention. It is hard for me to believe that this could be wr

ong or cruel when

both sides of the situation arc benefiting and not being harmed. It is experiences like this that

make

me feel lucky to attend a university that offers opportunities that will

help me learn and grow in ways I

never thought were possible.

Sincerely, A proud Cal Poly Pomona animal veterinary science undergradu

ate student

Catherine Pabst

And the winners are:


Megan Ducey-HardosI am a student at Cal Poly Pomona majoring in animal health science. Like you, throughout

my life, I have had a passion for helping animals. My passion has led me to pursue a career

in the animal health science field as a veterinary technologist. When I was in high school, I

had the same opinion you did about animal research. I thought it was horrible practice that

was torturing animals. After visiting our university research facility, I realized my current

opinion of this field was very inaccurate. The animals at these facilities were being humanely

treated and were given an enriched habitat. During my sophomore year of college my

mother was diagnosed with breast cancer. It is hard to see someone you love go through

the pain of having a debilitating and often fatal disease. After seeing my mother go through

the treatments and therapies, I realized that animal research right now is necessary to find

cures, treatments, and therapies for diseases. If it wasn’t for animal research, my mother

wouldn’t be here today. I hope for the day that we don’t have to use animals but right now,

animal research is needed for the testing of biomedical drugs and studies on certain

diseases. When I graduate, I want to pursue a career as a research animal manager

to promote the humane treatment of laboratory animals of all species and models.

Currently, there are several research projects being conducted at our University that

can only be performed in animals. Our IACUC always looks at alternatives and utilized the 3 R’s when reviewing a protocol. If the research can be performed in vitro, they won’t use laboratory animals. Two research projects can only use animals and they could discover the treatments and therapies for Huntington’s disease and GABA transport membrane related diseases. One research project uses Xenopus laevis to study GABA transport membranes. In this research, researchers surgically remove oocytes from female Xenopus laevis frogs. The oocytes are observed under a freeze-fracture and electron microscope. The goal is to see what does or doesn’t affect the GABA transport membrane. Pharmaceutical companies can use the information from this study to develop drugs for illnesses like Huntington’s and Parkinson’s disease. Oocytes of humans can’t be used for this study because they are too small to visualize and study. Frogs have bigger oocytes than any other animal. Frogs are not harmed in the research or tortured. These frog are anesthetized during the procedure and don’t feel any pain from the egg removal. These frogs receive a great degree of enrichment and a wide range of food.

The second research project that is performed at our University Research Facility utilizes mice for Huntington’s disease studies. In this research, we study the huntingtin protein in the cardiac and skeletal muscles of mice. R6/2 mice models that resemble patients with Huntington’s disease are utilized in this research. At around 12 weeks mice will display the first stages of the disease which is weight and muscle loss. The researchers care about the welfare and the pain intensity each mouse experiences. When mice display the first stages of the disease, they are humanely euthanized. Your organization believes euthanasia is necessary if animal is experience incurable pain or diseases. We also agree that our laboratory animals should be humanely euthanized and should not experience the later stages of Huntington’s disease. Once euthanized, the mouse’s cardiac and skeletal muscles samples are removed andobtained.Microelectrodes are used to inject electrical currents into mice muscle fibers. In this research, we found that this disease changes membrane potential in muscles and in addition reduces chloride channels. This research can help find treatments and therapies for

this debilitating disease.

Fiona LairThe world of animal research has come a long way since its first utilization. It is true,

in its early days, animal research did have cruel and unethical practices that were

not necessarily in the best interest of the animal or human participants. But we have

come a long way since the days where dogs, cats, mice, frogs, or any other animal

could be snatched up off the street and experimented on at the whim of the scientist.

Thanks to various and numerous interest groups, government agencies, and

environmental policies, animals in research are treated better than ever. Many

animals are even treated better than personal pets that people keep in their homes.

Research animals are provided with every need they could possibly have, unless that

need is what is being examined, at every hour of the day. Even with adoption processes,

people are still able to get a hold of any number of animals and there are no inspections

and officials coming to make sure those animals are being treated properly.

In research facilities throughout the United States, Cal Poly Pomona included, before a

researcher even can get money to look at catalogs in earnest to purchase animals, they

have to go through mountains of paperwork and get so many approvals, many would-be

animal researchers are likely deterred by the sheer amount of work it takes to get even a

mouse, let alone a more complicated and less readily accepted research animal. This

process is necessary not only to weed out those people who are not serious enough to put

in the effort it takes to get a research plan approved, but to ensure that every angle has

been considered for every possible scenario that could happen to any of the animals

during the course of the research.Researchers do not want to put their animals in any more pain then they absolutely have to.

They go through extraordinary precautions and protocols just to make sure that the animals

they work with have the highest quality of life they possibly can while still getting accurate

results. If an animal were to fall ill during the course of a study, an illness that was not

induced that is, they would be immediately treated as much as they could. All appropriate

anesthesia and analgesia protocols to alleviate discomfort and pain that may be necessary

to the study are used in order to make them more comfortable. Unfortunately, pain, illness,

and discomfort are a necessary part of the research process. If pain, illness, and discomfort

were not a part of everyday life for many people, and animals, many areas of research

would not be necessary.Animal testing is not all about poking and prodding around and seeing how things work for

the sake of curiosity. There has to be a justifiable need and sufficient desire from funding

sources for a problem to be solved in order for research proposals even to be considered. It

serves not only to help the biomedical community, but anatomy, physiology, and disease

mechanics as well. Without animal research, we would not know what we know now about

viruses and bacteria and how we treat and prevent them. We would not have vaccinations

or many treatments. With animals testing, tragedies like those that followed the release of

thalidomide, or even Agent Orange, can be avoided.The sacrifices that animals make in the name of science are ones that help every person

that is alive today, will be alive in the future, and people who are now deceased. It is true

that at one point in time animal testing was not the highly regulated and officiated process

that it is today, but times have changed. Animals in research are now respected and

regarded at integral parts of the research process, They are not abused or mistreated. All

the treatments that they receive are carefully reviewed and documented to ensure that they

are experiencing the best quality of life as possible. For most areas of research substitutions

are either nor practical or not possible. Animals provide an insight to the workings of the

body that artificial models simply can’t.

Established in 1981, FBR is the nation’s

oldest and largest organization devoted

to educating the public about the essential

role of biomedical research in the quest

for medical advancements, treatments

and cures for both humans and animals.

For more information, visit fbresearch.org.


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CANCER CURIOSITY:Gorbunova and Seluanovfocused on the biology ofcancer after noticing thatrates of the disease varywidely across a set of 15species of rodents thatincluded beavers (inset).


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A genetic twist in a rarely studied animal may have bigimplications for the fight against cancer in humans. by Jonathan Sherwood ‘04 (MA), ‘09S (MBA)

Phone calls at 3 a.m. rarely involve good news. Especially if the caller is a man toting a firearm. But to Vera Gorbunova, an associate professor ofbiology, the call that woke her was a welcome one. On the phone was a hunter fresh from the swamps of the Montezuma National Wildlife Refuge near Cayuga Lake, east of Rochester.

The hunter knew that the Rochester professor was on the lookout forhard-to-come-by rodents. And beavers fit the bill. Normally protectedfrom hunting and trapping in New York, the mostly nocturnal animalsare considered pests in the wildlife refuge because of the damage theycan do to the swamp’s white oak and birch trees.

Beavers were among the 15 rodents Gorbunova was studying toinvestigate a hunch she had about cancer.

Why, she had asked, can a squirrel livenearly two de-cades—well into thegolden years for a relatively smallmammal—and not show signs ofcancer? Yet mice, if they manage tolive past two years old, often succumbto the disease?

Is it possible, Gorbunova wondered,that some rodents have ways to protectagainst cancer that are completely unknown to humans? ➤

The MYSTERY

of an Antıcancer Mechanism


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This fall, Gorbunova may have found an intriguing answer inone of the stranger rodents on her list—the east African nakedmole rat, mice-like creatures that spend their livesunderground in highly social colonies. Naked mole rats canlive up to 28 years, the longest lifespan of any rodent, yet theyhave never been observed to develop cancer.

In a paper published in the Proceedings of the NationalAcademy of Sciences, Gorbunova reported that naked molerats seem to have a genetic ability to stop cells from replicatingif too many crowd together. And runaway cellular replication,she notes, is the very definition of cancer.

“Gorbunova has put her finger on the mechanism that gives adramatic cancer resistance to this rodent,” says John Sedivy, aprofessor of biology and medical science at Brown University.“Her work elegantly demonstrates the value of studying‘unusual’ animals because this mechanism simply does notappear to exist in mice.”

Searching for such anticancer mechanisms in animals that arenaturally long-lived has been the goal of Gorbunova and herhusband and long-time collaborator, Andrei Seluanov, anassistant professor of biology, since they arrived at theUniversity in 2004.

Attracted by Rochester’s unusually strong combination of bothmolecular and evolutionary biology, the pair set up a lab inHutchison Hall, where they now oversee a research team thatincludes a halfdozen graduate students and a half-dozenundergraduates.

One of only a few research groups across the country to studythe seemingly “cancer-proof” naked mole rats, the teamfocuses on the role of telomerase, an enzyme that plays a keyrole in cellular replication that has been an important vein ofcancer research over the past 25 years. (The team thatdiscovered the enzyme’s role in 1985 received the Nobel Prizein Medicine and Physiology in December.) Something like amolecular housekeeper, telomerase makes sure that the endsof chromosomes—brief sections of DNA called telomeres—stay intact. As each cell divides, its telomeres slowly shorten,eventually resulting in the death of the cell. Withouttelomerase, the telomeres would shorten much sooner. Iftelomerase were to act just right, cells conceivably couldreproduce forever.

For Gorbunova, as with most cancer researchers, studying thedisease traditionally has meant focusing on one of two models:mice or humans. A key difference between the two organismsis that in mice, telomerase is very active, allowing cells toreproduce quickly. In humans, telomerase is much less active.On the plus side, that means mice heal from injuries far fasterthan humans do.But there’s a downside—increased cancer riskas unwanted cells reproduce quickly and indefinitely.

Because telomerase allows cells to reproduce veryquickly, biologists had long assumed that thereason humans suppress the action of telomeraseand mice don’t is that mice live on average onlyabout two years. Their risk of getting cancer is lowbecause they’re not likely to live long enough toget the disease. But humans live for 80 years,

plenty of time to develop a few cells that will become cancerous.Gorbunova, however, didn’t accept that explanation. In tests ofseveral closely related rodents that varied greatly in lifespan, sheexplored whether longer-lived animals suppressed theirtelomerase more than shorter-lived ones.

“Selecting the rodents was easy, but getting the tissue samples from each of them was much, muchharder. Some of the rodents, like the beaver, are protected species. Some don’t exist in North America.Some weigh more than a hundred pounds. It’s not like you can order them out of a catalog. It took us

more than a year of calling and e-mailing all sorts of people to find all the rodents we needed.”


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“Selecting the rodents was easy, but getting the tissue samples fromeach of them was much, much harder,” says Gorbunova. “Some ofthe rodents, like the beaver, are protected species. Some don’t existin North America. Some weigh more than a hundred pounds. It’snot like you can order them out of a catalog. It took us more than ayear of calling and e-mailing all sorts of people to find all therodents we needed.”

The list included regular mice, squirrels, otters, and gerbils, as wellas more exotic animals such as giant capybara from SouthAmerica, chinchillas, and the naked mole rat.

As Gorbunova studied the tissue samples over the years, she was surprised to find no correlation betweenhow long a rodent lived and the action of itstelomerase. Some animals, such as the naked mole rat, lived nearly three decades yet expressed as muchtelomerase as a regular mouse.

Instead, another correlation came to light—body mass. Largeranimals, like humans and capybara, simply have more cells thatcan become cancerous, and so telomerase is suppressed to reducethe chance that any particular cell will set off a tumorous cascade.But what about the naked mole rat? Despite their long lives and thelarge numbers of naked mole rats under observation, there hasnever been a single recorded case of a mole rat contracting cancer,says Gorbunova.

Adding to the mystery is the fact that mole rats appear to age verylittle until the very end of their lives.

When Gorbunova and her team began investigating mole rat cells,they were surprised at how difficult it was to grow the cells in thelab. The cells simply refused to replicate once a certain numberoccupied a space. The mole rats seemed to be able to turn theprocess of replication off regardless of the action of telomerase.

Other cells, including human cells, also cease replication whentheir populations become too dense, but the mole rat cells werereaching their limit much earlier than those of other animals.Gorbunova and Seluanov have named the phenomenon “earlycontact inhibition.”

“Since cancer is basically runaway cell replication, we realized thatwhatever was doing this was probably the same thing that preventedcancer from ever getting started in the mole rats,” says Gorbunova.

That early contact inhibition was so pronounced that whenGorbunova’s team mutated cells to induce a tumor, the growth ofcells in the naked mole rats barely changed, whereas mouse cellsbecame fully cancerous.

“We think we’ve found the reason these mole rats don’t get cancer,and it’s a bit of a surprise,” says Gorbunova. “It’s very early tospeculate about the implications, but if the effect of early contactinhibition can be simulated in humans we might have a way to haltcancer before it starts.”

The key, according to Gorbunova, can be found in the action of a gene known as p16, which, in naked mole rats, triggers an early anticancer mechanism that appears to tell the cells to stop replicating. As in many animals, including humans, the molerats also have a gene called p27 that limits how many cells cancrowd together.

“In humans and mice, p16 doesn’t play a major role in contactinhibition, but in the naked mole rat p16 gets activated when thecells just begin crowding and arrests cell proliferation,” Gorbunovasays. “Cancer cells tend to find ways around p27, but mole rats have a double barrier that a cell must overcome before it can grow uncontrollably.

“We believe the additional layer of protection conferred by this two-tiered contact inhibition contributes to the remarkable tumorresistance of the naked mole rat,” says Gorbunova. Gorbunova andSeluanov plan to delve deeper into the mole rat’s genetics to see ifthe animals’ cancer resistance might be applicable to humans.

“The approach is promising as humans also have the p16 gene,but it plays a different role in anticancer protection,” saysGorbunova. “When we learn more about the differences betweenthe gene in humans and naked mole rats we may learn how toactivate the earlier protection in human cells.

“It’s also important to study other genes involved in cell-to-cellcontact in order to understand how early contact inhibition canserve to cure or even to prevent human cancer.”

Jonathan Sherwood ‘04 (MA), ‘09S (MBA) is a senior science writer forUniversity Communications.

ANTICANCER KEY?Despite a long lifespan,

naked mole rats (this page) have neverbeen observed to havecancer, a disease thatoccurs in varying ratesin squirrels, capybaras,

otters (opposite), and other rodents,

according toGorbunova.

AFTER 4 YEARS OF MEDICAL SCHOOLAND 2 YEARS of training to be a brainsurgeon, Kevin Tracey had learned to controlthe emotions he felt for his patients. Buteverything changed on May 3, 1985, whenhe met Janice.

As part of his training, Tracey was working inthe emergency room at Cornell University’shospital in New York City. At 6:45 p.m.,paramedics brought in an 11montholdfemale with severe burns. Tracey examinedher and noted: burns on 75 percent of herbody, no broken bones, no other injuries.

As he focused on his patient’s needs, Traceyheld back tears. Janice, who arrived wrappedin a teddy bear blanket, now writhed andsobbed on a metal gurney. Her oncesoft skin was seared and peeling from her arms,legs and back. She had a 25 percent chanceof surviving.

Just an hour before, Janice had beengiggling on the kitchen floor while hergrandma cooked spaghetti. When thegrandma turned to drain a 10quart pot ofboiling noodles, she tripped and spilled the212degree water all over the baby. Sweetchuckles turned to inconsolable screams.

For the next 3 weeks, Tracey and otherswatched Janice ride a rollercoaster ofrecoveries and setbacks.

First, she went into a 5hour surgery, whereTracey and his colleagues removed scaldedskin and replaced it with thin layers shavedfrom Janice’s bottom—an area that had beenprotected by her diaper. Everyone—her familyand her medical team—sighed with reliefonce Janice opened her eyes and smiled.

The next night, though, her blood pressuredropped dangerously low, starving her brain, kidneys, lungs and other vital systemsof muchneeded oxygen. Her body was inseptic shock. Tracey immediately pumpedfluids and drugs into her veins to raise herblood pressure and prevent permanent tissue damage.

Despite these measures, Janice drifted into acoma. The next day, her organs beganworking again and she awoke.

But then she spiked a fever and her bodyswelled. Her kidneys—and soon her liver andother organs—stopped working. Janice wasexperiencing widespread inflammation, alifethreatening condition called sepsis. Asbefore, Tracey’s infusion brought her back to life.

–Photo by ADA

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For Janice:Legacy of a Short Life BY EMILY CARLSON


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On May 28, Tracey celebrated with herfamily as Janice turned 1 year old. Insteadof machines and monitors, balloons andstreamers now filled her room.

But the very next day, Janice’s heartstopped. This time, no medical feat could bring her back.

A New PathStunned by her sudden, unexpecteddeath, Tracey set out to understand whatcaused it and to prevent the same thingfrom happening to anyone else.

During the next 2 years, he put hisneurosurgery career on hold and focusedon medical research.

“Most doctors have a patient who affectstheir lives, who they really wish they couldhave done something more for,” saysTracey. “Janice is my patient, and she hasa lot to do with the path I’ve been on ever since.”

Tracey was particularly curious to figureout why Janice’s blood pressure haddropped so low. He knew her conditionstemmed from septic shock, when thebody’s immune system reacts violently to abacterial infection. But he had run all sortsof blood tests to locate an infection andhad never found one.

If there was no infection, then whatcaused her immune system to rage out ofcontrol? The answer could overturncenturies of thinking about what makes us sick.

Defense, Defense, DefenseThe immune system is the body’s naturaldefense against viruses, bacteria andother invaders. Its army is made up ofmore than 15 different types of whiteblood cells that produce molecules todefend and protect our bodies.

Some white blood cells produce proteinscalled antibodies that bind to particularinvaders and disable their actions. Othersmake proteins called cytokines that, whenreleased into an infected area, help healwounds and repair damaged tissue.

You can tell when your immune system isworking because you get a fever, swollenlymph nodes in your neck, redness arounda wound or even a rash or hives. Theseare the hallmarks of inflammation, animmune response designed to kill foreigninvaders.

But for Janice, it had a different effect. Herimmune system had begun to destroy thevery thing it was supposed to protect.Tracey set out to discover why and how.

Cytokine CuresTracey’s mentor and others had justidentified a type of cytokine abbreviatedTNF. Their research in mice suggested thatTNF might play a role in infection.

Wondering whether TNF had beeninvolved in Janice’s case, Tracey injectedrats with the cytokine. Almost immediately,their blood pressure plummeted and theywent into septic shock.

Like Janice, the rats had a high whiteblood cell count, suggesting infection. Butagain, there was no bacterial infection—just an excess of TNF.

These experiments convinced Tracey thattoo much TNF can cause septic shock inrats. He further reasoned that, since ratsand people are biologically similar, TNFprobably does the same thing in humans.

Committed to finding a better treatmentfor patients, Tracey and his team createdan antibody that could latch onto andimmobilize TNF. It worked in laboratorydishes, but could it soak up excess TNF inliving organisms? Could it stop septicshock, preventing harm to healthy organsand tissues?

To find out, the scientists tested theantibody in baboons whose bloodstreamswere filled with live bacteria, a conditionknown to cause septic shock.

Bingo! The antibody protected the animals.

While the antiTNF antibody was neverdeveloped into a drug, other scientistsbuilt on Tracey’s work and soondeveloped antiTNF medicines that are

now used to treat inflammationtriggeredarthritis in millions of people.

Ulf Andersson, a doctor in Sweden, sayshe had been waiting decades for amedicine like this. He treats kids withinflammatory conditions like juvenilearthritis. When they come to him, many ofhis patients ache from inflamed joints intheir arms and legs, making it difficult forthem to move and even grow. The drugcan dramatically improve the quality oftheir lives, Andersson reports.

“I have children who have been inwheelchairs for years who now play soccer again.”

Compassion CuresAndersson describes Tracey not only as a scientific collaborator but also as aclose friend.

“I admire his human character as much ashis genius,” Andersson says.

After Andersson’s wife died, Tracey flewfrom New York to Sweden to cheer up hisfriend. In the dead of winter, they put onspecial long skates and glided acrossparts of the Baltic Sea. Tracey’s comfortand support, says Andersson, brought himback to life. ➤

White blood cells, part of the immune system, protect us from viruses, bacteria and other invaders.

– IMAGE COURTESY JIM EHRMAN, DIGITALMICROSCOPY FACILITY, MOUNT ALLISON UNIVERSITY


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Andersson returns the favor twice a year, spending a week withTracey, his wife and their four daughters.

“It is really amazing how he finds the time to perform firstclassscience and manage to be the best father and husband I haveever seen,” says Andersson, who admits to copying some of hisfriend’s family skills with his own children.

Tracey attends each of his daughters’ soccer games and says hewould attend every single practice if he could. He wants them tohave fun, follow their passions and gain confidence in all thatthey do.

“Kevin is at his peak performance when he runs along thesidelines providing advice about how to play,” says Andersson.“As an American, his understanding of soccer equals his lack ofskill on the skates,” he jokes.

“We all like being on the water,” Tracey says. “A couple of hoursfeel like 2 days off.”

The Octopus and the O.R.Tracey decided on a career path when he was just 5 years old,right after his mother died. Sitting on his grandpa’s lap, he askedwhat had happened to her and why. His grandpa said she had atumor that, like an octopus, had spread its tentacles throughouther brain, making it impossible to remove. Right then, Tracey saidhe wanted to be a scientist so other kids didn’t have to suffer like him.

He also liked caring for people, which steered him towardmedicine.

Eventually, he decided to become a neurosurgeon—a specialtythat allowed him to split his time between the operating roomand the research lab. This dual career, he says, made him both abetter doctor and a better scientist.

“Being a doctor is gratifying in part because of the experiencethat comes from helping one person at a time,” he says. “But Iwas always excited about the possibility of discovering somethingthat could help many, many people.”

Also, he says, “Doing science is just addictive. It is more like ahobby than a job.”

“For every 100 questions, there are another 100 questions. Thechurning of these questions and the development of new ideasare what drive scientific progress.”

Surprise in the LabIn 2000, Tracey left his medical practice to devote all his energyto the lab. He turned his attention again to creating an antiTNFdrug.

He and his team developed a chemical called CNI1493. Thiscompound, which could switch off TNF production, had thepotential to treat cytokinerelated disorders from sepsis andarthritis to stroke and digestive diseases.

Her immune system had begun to destroy the very thing it was supposed to protect.

For Tracey and coworkers, lively discussions often lead to new questions to explore and answer.

– Photo by ADAM COOPER


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Tracey knew the substance targeted immune cells. What surprisedhim was that it had an even more powerful effect on brain cells.

That’s weird, Tracey thought. Why would something designed totarget the immune system have an effect in the nervous system?

He discovered that CNI1493 tells brain cells to activate aparticular nerve, called the vagus nerve. Once activated, the nerveturns down TNF production.

The WandererWe know that the nervous system controls many importantfunctions —from moving muscles to forming thoughts. But untilquite recently, scientists believed that the immune systemfunctioned independently.

Now it’s clear that the brain and other parts of the nervous system—probably a whole lot of nerves, Tracey suspects—help direct ourimmune responses.Tracey focused on the vagus nerve. This nerve,which means “wandering” in Latin, regulates our heart rate,digestion and other essential functions.

It meanders from the brain stem, across the neck and chest, down into the abdomen and ends up in our internal organs,including the spleen. Like the appendix, the spleen is an organ you probably think little about. But it’s actually a major player inthe immune system.

“Most of our circulating [white blood] cells pass through thespleen every 5 minutes,” says Tracey.

Thebraincommunicateswiththese white blood cellsvia the vagus.Electrical signals fromthebrainzip down the nerveand triggeritsendingsto release amolecule called acetylcholine into thespleen. When acetylcholine binds to special receptors on whiteblood cells, the cells stop making TNF. Less TNF means lessinflammation.

Sketching Out an IdeaSince TNF causes septic shock, Tracey wondered how the brainlimits production of the cytokine. To work out the answer, heturned to a trick he often uses to capture his thoughts: sketching.

When he first examined Janice, he diagrammed her burns andtheir severity. When he planned to build his daughters a twostoryplayhouse, he drew a blueprint on the back of a grocery bag.When he talked to middle school students about being a brainsurgeon, he went to the blackboard and drew a drill with a springto help the kids figure out how to bore through the skull withoutnicking the brain.

Now using a whiteboard in his lab, Tracey sketched the brain, thespleen and the vagus nerve running between them.

The next day, he set up an experiment to see if, by stimulating thevagus nerve, he could dampen TNF production. He used an

electrical device to activate the vagus nerve of rats. Withinseconds, the animals produced less TNF.

Since then, he has shown that in animals, stimulating the vagusnerve can block arthritis, sepsis, shock, heart failure andinflammation of the colon and pancreas.

“The groundwork is being laid to try some of these approaches inhumans, perhaps within the next year,” he says.

Legacy of LifeWhen Janice died, Tracey didn’t know exactly what happened or why.

“I had no good explanation for what the molecules in her bodywere doing,” he says. “I know a tremendous amount more now.”

He suspects that the baby’s injury impaired her nervous system,which not only led to an overproduction of TNF but also to herheart failure. He also thinks that too much HMGB1— anothertype of cytokine discovered in his lab—likely triggered her sepsis.

More than 25 years later, Janice’s story still influences Tracey everyday.

“I tell everyone who will listen never to take anything out of themicrowave with a baby in their arms,” he says. “Never put hotfood on a tablecloth that a child could pull down.”

He created an invisible triangle that fills the space between his

kitchen sink, stove and refrigerator. From an early age, his girls—now 7 to 15 years old—knew not to step inside it whenunsupervised.

He even wrote a book called Fatal Sequence: The Killer Within.The title refers to septic shock caused by the immune system. The book describes Janice’s life, how she changed Tracey’s lifeand what he and others have learned—and applied to helppatients—because of her.

Although she didn’t live to see it, her short life had a big impact, says Tracey. “The people who receive drugs based either directly or indirectly onknowledge that came [because of] her—they’re her legacy.”

For every 100 questions, there are another 100 questions.

Her short life had a big impact.

Tracey’s book describesJanice’s life and what heand others have learnedabout septic shock.

– Photo courtesy of THEDANA FOUNDATION

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